Chromosome Res (2023) 31:18
https://doi.org/10.1007/s10577-023-09727-7
REVIEW
Twenty years of merotelic kinetochore attachments:
a historical perspective
Daniela Cimini
Received: 1 May 2023 / Revised: 20 June 2023 / Accepted: 8 July 2023 / Published online: 19 July 2023
© The Author(s), under exclusive licence to Springer Nature B.V. 2023
Abstract Micronuclei, small DNA-containing
structures separate from the main nucleus, were
used for decades as an indicator of genotoxic dam-
age. Micronuclei containing whole chromosomes
were considered a biomarker of aneuploidy and were
believed to form, upon mitotic exit, from chromo-
somes that lagged behind in anaphase as all other
chromosomes segregated to the poles of the mitotic
spindle. However, the mechanism responsible for
inducing anaphase lagging chromosomes remained
unknown until just over twenty years ago. Here, I
summarize what preceded and what followed this dis-
covery, highlighting some of the open questions and
opportunities for future investigation.
Keywords lagging chromosome · merotelic ·
micronuclei · chromosome segregation · mitosis
Introduction
During mitosis, the sister chromatids of the repli-
cated chromosomes must separate and segregate to
opposite poles, and this process must be executed
Responsible Editor: Stefano Santaguida
D. Cimini (*)
Department of Biological Sciences and Fralin Life
Sciences Institute, Virginia Tech, Blacksburg, VA 24061,
USA
e-mail: cimini@vt.edu
accurately to ensure that each daughter inherits a
complete copy of the genome. A variety of mitotic
defects can lead to chromosome missegregation and
result in genetic imbalances in the daughter cells
[reviewed in (Baudoin and Cimini 2018b)]. Many of
these errors are prevented by a biochemical signaling
pathway, known as the spindle assembly checkpoint
(or SAC), that monitors chromosome attachment to
the mitotic spindle (Lara-Gonzalez et al. 2021). Ana-
phase lagging chromosomes, however, are caused
by a defect that can escape SAC detection and there-
fore represent a major threat to mitotic fidelity. Upon
mitotic exit, anaphase lagging chromosomes often
form micronuclei in the nascent daughter cell (Cimini
et al. 2002; He et al. 2019) and micronuclei were
shown to cause a variety of adverse effects (Krupina
et al. 2021). Although other chromosome segregation
defects can be observed in cancer cells and tumor tis-
sues (Gisselsson et al. 2004; Stewenius et al. 2007;
Tucker et al. 2023), anaphase lagging chromosomes
were shown to be prevalent in chromosomally unsta-
ble cancer cells, mouse tumor models, and certain
human tumors (Bakhoum et al. 2011; Bakhoum et al.
2015; Bakhoum et al. 2014; Thompson and Comp-
ton 2008; Venkatesan et al. 2021; Zaki et al. 2014).
Similarly, micronuclei are often found in cancer cells
and tumor tissues (Gisselsson et al. 2001; Streffer
et al. 1985), and are acknowledged to be an impor-
tant biomarker associated with increased risk of dis-
ease, including cancer (Bonassi et al. 2011a, b). The
mechanism responsible for the formation of lagging
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chromosomes was discovered just over twenty years
ago (Cimini et al. 2001). That discovery became the
seed for a series of new research questions, ranging
from the correction mechanisms, to the mechanisms
that increase these errors in cancer cells, to the biol-
ogy of micronuclei, and more. In this review, I sum-
marize questions, knowledge, and ideas that preceded
the identification of the mechanism underlying ana-
phase lagging chromosomes. Then, I briefly summa-
rize all the findings that followed that discovery and
are still a fertile ground of investigation for research-
ers in the chromosome segregation, aneuploidy, and
cancer biology communities.
Anaphase lagging chromosomes
and kinetochore‑positive micronuclei
as biomarkers of aneuploidy
An anaphase lagging chromosome is defined as a
chromosome that lags behind during anaphase, when
all other chromosomes segregate to the spindle poles,
and it can form a micronucleus upon mitotic exit
(Fig. 1B). A micronucleus, as the name suggests, is
a small structure consisting of DNA encapsulated by
a nuclear envelope, which is separated from the main
nucleus. By the early 1990s, the cellular toxicology
community viewed anaphase lagging chromosomes
as precursors of certain micronuclei, whose occur-
rence was considered to be a biomarker of aneuploidy
(Antoccia et al. 1991; Bonatti et al. 1992; Degrassi
and Tanzarella 1988; Thomson and Perry 1988).
This was partly based on detailed cytology studies
indicating that missegregation of chromosomes or
chromatids and subsequent micronucleus formation
could lead to aneuploidy in the cell progeny (Gus-
tavino et al. 1994; Rizzoni et al. 1989). Detecting and
quantifying micronuclei is simple and fast and there-
fore they have been used as a measure of genotoxic
damage for decades (Pincu et al. 1985) in a stand-
ardized assay named the in vitro micronucleus assay
(Fenech 2000). However, micronuclei can form as

Fig. 1  Diagram showing

formation of micronuclei
upon mitotic exit. Chromo-
somes/DNA are depicted
in blue, kinetochores in
orange. A. Formation of a
kinetochore-negative micro-
nucleus from a lagging
chromosome fragment. B.
Formation of a kinetochore-
positive micronucleus
from an anaphase lagging
chromosome. C. Formation
of a kinetochore-positive
micronucleus in a binucle-
ate cell in the cytokinesis-
block assay

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a result of either missegregation of whole chromo-
somes (Fig. 1B) or acentric chromosome fragments
(Fig. 1A) that cannot interact with the mitotic spin-
dle, and the simple visualization of micronuclei does
not provide information on their origin. In the late
1980s, the genetic toxicology community recognized
the need to identify genotoxic agents that affected
cells and organisms in ways other than by damaging
their DNA or inducing gene mutations. Specifically,
it had become apparent that certain chemical agents
do not cause DNA damage (i.e., are not clastogenic),
but can cause aneuploidy (i.e., are aneugenic), which
can severely impair cell and organismal physiology.
At that time, the method of choice for assessing ane-
uploidy was counting chromosomes in chromosome
spreads. However, such approach is time consuming
and has limited statistical power, given that typically
only ~ 50 chromosome spreads are analyzed per sam-
ple. Therefore, there was a need to develop an assay
that allowed to quantify aneuploidy quickly and eas-
ily. The in vitro micronucleus assay appeared to
have the right requisites, but only if strategies could
be devised to distinguish micronuclei that contained
entire chromosomes (from aneugenic effects) from
those containing chromosome fragments (from clas-
togenic effects). To this end, attempts were made to
discriminate micronuclei based on their size and/or
their DNA content, assuming that micronuclei that
were larger and/or contained more DNA, were more
likely to contain entire chromosomes compared to
smaller micronuclei (Hogstedt and Karlsson 1985;
Pincu et al. 1985; Yamamoto and Kikuchi 1980). This
idea was justified, given that micronuclei containing
larger chromosomes were recently reported to have
a larger area (Mammel et al. 2022). However, these
approaches are laborious and difficult to standardize
and therefore were never adopted as routine methods
in the genetic toxicology field. The discovery of anti-
bodies from scleroderma patients (CREST) that local-
ized to the kinetochore (Moroi et al. 1980) enabled
the discrimination between micronuclei containing
entire chromosomes (CREST-positive micronuclei)
and micronuclei containing chromosome fragments
(CREST-negative micronuclei) (Fig. 1A-B). A num-
ber of studies focused on assessing the ability of
such approach to predict the aneugenic potential of
physical and chemical agents (Antoccia et al. 1991;
Fenech and Morley 1989; Hennig et al. 1988; Schuler
et al. 1997). As opposed to the detection of anaphase
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lagging chromosomes or counting chromosomes in
chromosome spreads, this approach could be used to
quantify micronuclei in thousands of interphase cells
in tens of minutes (Fenech 2000). A variation of this
approach was subsequently developed to selectively
identify micronuclei in cells that were proliferative
and underwent cell division during or after exposure.
This method was based on the idea that aneuploidy
arising in proliferative cells would affect the cell pop-
ulation, and hence organismal health; moreover, this
approach would prevent micronuclei already present
in the cell population from confounding the data. In
this approach, named the cytokinesis-block assay,
cells were treated, during or after exposure, with
cytochalasin (an inhibitor of actin polymerization) for
a time window corresponding approximately to cell
cycle duration prior to fixation and analysis. All the
cells undergoing cell division during the cytochalasin
treatment would be unable to complete cytokinesis
and would therefore appear as binucleate cells; analy-
sis of micronuclei could then focus specifically on the
binucleate cell population (Fig. 1C). The combination
of the cytochalasin treatment with kinetochore stain-
ing was viewed as a powerful method in the genetic
toxicology community (Fenech 2000), although com-
parison of rates of anaphase lagging chromosomes
vs. CREST-positive micronuclei in binucleate cells
revealed that this method underestimated the num-
ber of anaphase lagging chromosomes (Cimini et al.
1999).
Although the genetic toxicology community
broadly used centromere/kinetochore-positive micro-
nuclei as a readout of aneuploidy and overall agreed
that such micronuclei originated from anaphase lag-
ging chromosomes, the mechanism causing lagging
chromosomes remained unknown.
Merotelic kinetochore attachment: the underlying
cause of anaphase lagging chromosomes
Because the genetic toxicology community was not
accustomed to thinking about cellular mechanisms
and the cell biology community was not focused on
aneuploidy and its consequences, the cellular mech-
anism causing chromosomes to lag behind during
anaphase was unknown until just over 20 years ago.
Nevertheless, some ideas were shared in informal dis-
cussions and included overriding of the SAC in the
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presence of unattached chromosomes/kinetochores,
accidental detachment of microtubules from kineto-
chores during anaphase, and failure of sister chroma-
tids to separate (which would lead to the formation
of micronuclei with two kinetochores) as the poten-
tial causes of anaphase lagging chromosomes. The
idea that anaphase lagging chromosomes may result
from a “leaky” SAC was perhaps influenced by the
term used for chromosomes that, in prometaphase
cells, were positioned close the spindle poles. These
chromosomes were often referred to as “lagging
chromosomes” and perhaps this terminology led to
the idea that these unaligned prometaphase chromo-
somes failed to align at the metaphase plate prior to
anaphase onset and were the precursors of anaphase
lagging chromosomes. We now prefer to refer to such
prometaphase chromosomes as “unaligned” chromo-
somes to differentiate them from the anaphase lagging
chromosomes (Baudoin and Cimini 2018b). It was
also shown that unaligned chromosomes are not seen
at anaphase onset in cells with anaphase lagging chro-
mosomes and that cells with anaphase lagging chro-
mosomes do not experience a pre-anaphase mitotic
delay (Cimini et al. 2002), which typically occurs
when unaligned chromosomes are present (Rieder
et al. 1994, 1995). These observations definitely dis-
missed the idea of anaphase onset in the presence
of unattached kinetochores as the underlying cause
of anaphase lagging chromosomes. The accidental
detachment of microtubules from the kinetochore was
also not plausible, given that the stability of micro-
tubules had been shown to dramatically increase in
anaphase compared to metaphase (Zhai et al. 1995)
and we now know that microtubule pulling forces sta-
bilize kinetochore-microtubule attachments (Akiyoshi
et al. 2010), thus making detachment in anaphase
highly unlikely. Finally, failure of sister chromatids
to separate was also very unlikely, given that staining
with individual centromeric probes showed micronu-
clei contained individual centromeres and anaphase
lagging chromosomes consisted of individual daugh-
ter chromosomes and not paired chromatids (Cata-
lan et al. 2000; Cimini et al. 1997, 1999; Falck et al.
2002).
Detailed analysis of chromosomes/kinetochores
and microtubules revealed that the underlying cause
of anaphase lagging chromosomes was merotelic
kinetochore attachment (Fig. 2), a type of kine-
tochore-microtubule attachment in which a single
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kinetochore binds microtubules from both spindle
poles instead of just one (Cimini et al. 2001). This
type of mis-attachment had previously been reported
in cold- or drug-treated crane-fly spermatocytes,
where the term merotelic was first used (Janicke and
LaFountain 1984; Ladrach and LaFountain 1986),
and in cells undergoing cell division with individual
chromatids or chromosome fragments containing
single kinetochores (Khodjakov et al. 1997; Wise
and Brinkley 1997; Yu and Dawe 2000). The obser-
vation that merotelic kinetochore attachment could
explain the origin of anaphase lagging chromosomes
in mitotic cells provided a possible mechanism for the
aneuploidy and chromosomal instability (CIN) fre-
quently observed in cancer cells (Bolhaqueiro et al.
2019; Lengauer et al. 1997; Nicholson and Cimini
2013; Rajagopalan and Lengauer 2004; Weaver and
Cleveland 2006). In support of this idea, subsequent
studies showed that most cancer cells display high
rates of anaphase lagging chromosomes (Bakhoum
et al. 2014, Ganem et al. 2009, Silkworth et al. 2009,
Thompson and Compton 2008) and lagging chromo-
somes were also observed in tumor tissue samples
(Bakhoum et al. 2011; Zaki et al. 2014), although
this may not be the case for all tumor types (Gissels-
son et al. 2004; Stewenius et al. 2007; Tucker et al.
2023). Merotelic kinetochore attachment was fur-
ther shown to be a key mechanism of aneuploidy in
human pluripotent stem cells (Deng et al. 2023) and
in mammalian oocytes, where it can promote chromo-
some missegregation in both meiosis I and II (Cheng
et al. 2017, Holubcova et al. 2015; Kouznetsova et al.
2014; Shomper et al. 2014, Zielinska et al. 2015),
although other mechanisms of chromosome misseg-
regation have also been described in meiotic cells.
For instance, one study identified lack of kinetochore-
microtubule attachment as a major cause of lag-
ging chromosome formation and aneuploidy in aged
mouse oocytes (Mihajlovic et al. 2021).
Overall, there is currently strong agreement that
merotelically attached lagging chromosomes repre-
sent a major source of aneuploidy and chromosomal
instability in normal and cancer cells.
How do merotelic attachments arise?
Establishment of kinetochore-microtubule attach-
ments is a stochastic process and therefore different
Chromosome Res (2023) 31:18
Fig. 2  Fate of anaphase lagging chromosomes depends on the
relative size of the microtubule bundles connecting the mero-
telic kinetochore to the two spindle poles. A. Example of live
cell (top) and diagram (middle) of a “balanced” merotelically
attached anaphase lagging chromosome. In the images (taken
two minutes apart), the lagging kinetochore can be seen per-
sisting at the cell equator as all the other chromosomes move
poleward. As a result of this, a whole chromosome-contain-
ing micronucleus is formed in one of the daughter cells (bot-
tom). B. Example of live cell (top) and diagram (middle) of an
“unbalanced” merotelically attached anaphase lagging chromo-
some. In the images (taken three minutes apart), two meroteli-
types of attachments can be established in early mito-
sis [reviewed in (Gregan et al. 2011; Sarangapani and
Asbury 2014)]. Merotelic kinetochore attachments
form often during the early stages of mitosis in nor-
mal bipolar cells, but they decrease as mitosis pro-
gresses (Cimini et al. 2003). Ultimately, chromosome
mis-segregation due to merotelic kinetochore attach-
ment will depend on the overall balance between
rates of formation of such mis-attachments and the
rates at which they are corrected. The rates of mero-
telic attachment formation were shown to increase in
several contexts, mostly having to do with either kine-
tochore structure or spindle geometry. For instance,
Page 5 of 15 18
cally attached kinetochores can be seen. However, because of
the different size of the two microtubule bundles connecting
the merotelic kinetochores to the two spindle poles, the chro-
mosomes’ position is shifted close to the group of normally
segregating chromosomes on the right. As a result of this, the
chromosome(s) will be included in the main nucleus in one
of the daughter cells (bottom). In the images, kinetochores
are in green and microtubules are red. In the diagrams, chro-
mosomes/DNA are depicted in blue, kinetochores in orange.
Live cell images are modified from Cimini et al. 2004 (A) and
Cimini et al. 2006 (B)
certain fission yeast mutants are believed to form high
rates of merotelic attachments because of disorgan-
ized kinetochore structure (Gregan et al. 2011, 2007;
Pidoux et al. 2000; Rumpf et al. 2010). Similarly, a
C. elegans mutant in which the kinetochore is twisted
around the chromatin forms high rates of merotelic
attachments (Stear and Roth 2002). Finally, in Indian
muntjac cells, whose chromosomes possess kineto-
chores of varying sizes, the probability of merotelic
attachment increases with kinetochore size (Baudoin
and Cimini 2018a, Drpic et al. 2018).
Spindle multipolarity is one spindle geom-
etry defect that was shown to increase the rates of
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merotelic kinetochore attachments. This happens
because within a multipolar spindle it is easier for a
single kinetochore to face more than one spindle pole
than it would be within a bipolar spindle (Ganem
et al. 2009; Paul et al. 2009; Silkworth and Cimini
2012, Silkworth et al. 2009) and indeed it was shown
that increasing numbers of spindle poles correlate
with increasing numbers of merotelic attachments
(Paul et al. 2009). This can explain earlier observa-
tions that lagging chromosomes are very commonly
observed in multipolar anaphases (Sluder et al.
1997) and in cells recovering from a nocodazole-
induced mitotic arrest (Cimini et al. 2001, 2003,
1999; Ladrach and LaFountain 1986). Indeed, in
cells recovering from a nocodazole arrest, microtu-
bules assemble from the two centrosomes as well as
from multiple ectopic microtubule organizing cent-
ers (MTOCs) (Cimini et al. 2003). However, these
ectopic MTOCs eventually cluster with the two cen-
trosomes to form a bipolar spindle (Cimini et al.
2003). The transient multipolarity state is sufficient
to enhance the rates of merotelic kinetochore attach-
ment and hence the rates of anaphase lagging chro-
mosomes (Ganem et al. 2009; Silkworth and Cimini
2012, Silkworth et al. 2009). Notably, this transient
multipolarity followed by centrosome clustering is
believed to play an important role in promoting chro-
mosome missegregation in cancer cells with super-
numerary centrosomes (Brinkley 2001; Ganem et al.
2009, Silkworth et al. 2009). Increased rates of mero-
telic attachment formation are also observed when the
centrosomes/spindle poles in mitotic cells do not sep-
arate prior to nuclear envelope breakdown (Kaseda
et al. 2012; Nam and van Deursen 2014; Silkworth
and Cimini 2012; Silkworth et al. 2012). This hap-
pens because, if the spindle poles are in close prox-
imity while kinetochore-microtubule attachments are
established, the chance of a single kinetochore to bind
microtubules from both spindle poles is higher than
it would be if the two spindle poles were at diamet-
rically opposed positions around the chromosomes
(Silkworth and Cimini 2012; Silkworth et al. 2012).
Because merotelic kinetochores are attached to
microtubules, the SAC is silenced and therefore
higher frequencies of merotelic attachments invari-
ably translate into higher frequencies of anaphase
lagging chromosomes. Nevertheless, correction
mechanisms exist in both prometaphase/metaphase
and anaphase to reduce the rates of chromosome
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missegregation from merotelic attachment. The next
section will focus on such mechanisms.
Correction of merotelic kinetochore attachments
throughout mitosis
Merotelic kinetochore attachments are very common
in early mitosis, but their numbers decrease as mitosis
progresses (Cimini et al. 2003, 2004). The Aurora B
kinase, which was already known to be essential for
chromosome bi-orientation in budding yeast (Biggins
et al. 1999; Tanaka et al. 2002), has been described
as a major player in the merotelic error correction
that occurs in prometaphase and metaphase (Cimini
et al. 2006) thanks to its ability to phosphorylate
kinetochore substrates, specifically the Hec1 subu-
nit of the Ndc80 complex, and cause microtubule
detachment (Cheeseman et al. 2006; Ciferri et al.
2008; DeLuca et al. 2006). Detachment of microtu-
bules from the kinetochore provides the opportunity
for loss of attachments from the incorrect pole and
formation of new attachments to microtubules from
the correct pole. Importantly, measurements of kine-
tochore-microtubule turnover showed that cells with
high rates of anaphase lagging chromosomes have
more stable kinetochore-microtubule attachments
in either prometaphase, metaphase, or both (Bak-
houm et al. 2009a). The exact mechanism by which
Aurora B kinase destabilizes kinetochore-microtubule
attachments is still debated, with large part of the
debate revolving around whether the pool of Aurora
B localized at the centromere or a pool localized at
the kinetochore is responsible for the phosphorylation
of Hec1 required to detach microtubules [for a review
on this topic see (Broad and DeLuca 2020)]. The cen-
tromere-localized kinesin-13s MCAK and Kif2b were
also shown to play a key role in the correction of kine-
tochore-microtubule mis-attachments, as their sup-
pression results in reduced turnover of kinetochore-
microtubules and increased rates of anaphase lagging
chromosomes (Bakhoum et al. 2009b; Kline-Smith
et al. 2004; Maney et al. 1998). Interestingly, Auora
B-dependent phosphorylation was also shown to
modulate MCAK activity (Andrews et al. 2004; Lan
et al. 2004; Zhang et al. 2007). Importantly, potenti-
ating MCAK using a small molecule was shown to
reduce the rates of anaphase lagging chromosomes
in cancer cells, suggesting that this missegregation
Chromosome Res (2023) 31:18
errors could be a druggable target, although the can-
cer cells quickly adapted and reestablished their
pre-treatment rates of lagging chromosomes (Orr
et al. 2016). A recent study has also suggested that
an interplay between Aurora B kinase activity and
mechanical tension specifically contributes to correc-
tion of merotelic kinetochore attachments as opposed
to kinetochore-microtubule mis-attachments lacking
tension (Chen et al. 2021). This model is somewhat
inconsistent with in vitro data showing that tension
can suppress Aurora B-mediated detachment of kine-
tochore-bound microtubules (de Regt et al. 2022), but
this may be because the interplay between tension
and Aurora B influences amphitelic and merotelic
attachments differently. Indeed, the Aurora B-medi-
ated correction of merotelic attachments may specifi-
cally depend on the deformation of merotelic kineto-
chores in metaphase and on the enrichment of Aurora
B at those sites (Cimini et al. 2004, 2006; Knowlton
et al. 2006; Trivedi et al. 2019). Finally, there is evi-
dence that, although a defective SAC may not directly
cause anaphase lagging chromosomes, premature
SAC silencing reduces the time window for the pre-
anaphase correction of merotelic attachments, leading
to increased rates of anaphase lagging chromosomes
(Cimini et al. 2003).
Merotelic kinetochore attachments do not sus-
tain SAC signaling the way unattached kinetochores
do [for a review on the SAC see (Lara-Gonzalez
et al. 2021)]. Therefore, although many merotelic
attachments can be corrected via the mechanisms
mentioned above, some can persist into anaphase.
Early studies showed that merotelic kinetochore
attachments that persist into anaphase can lead to
different outcomes. In some cases, merotelically
attached lagging chromosomes remain positioned at
the spindle equator, whereas in other cases they are
shifted toward one of the two groups of segregating
chromosomes (Cimini et al. 2004; Thompson and
Compton 2011). Anaphase lagging chromosomes
that remain close to the spindle equator, invariably
form micronuclei (Cimini et al. 2002), whereas
those that are shifted towards one of the two spin-
dle poles can be incorporated into the main nucleus
(Thompson and Compton 2011). The differential
positioning of lagging chromosomes during ana-
phase was shown to depend on the relative number
of microtubules emanating from the kinetochore of
the lagging chromosome to the two opposite poles
Page 7 of 15 18
(Cimini et al. 2004; Thompson and Compton 2011)
(Fig. 2). Specifically, if the lagging chromosome is
bound to microtubule bundles of similar thickness
(i.e., containing similar numbers of microtubules;
Fig. 2A), then the chromosome will remain close to
the cell equator (Cimini et al. 2004), whereas if the
two microtubule bundles are of different thickness
(as measured by fluorescence intensity; Fig. 2B),
then the chromosome will be shifted to one side
(Cimini et al. 2004), typically sufficiently close to
one of the two groups of segregating chromosomes
to avoid missegregation. Initially, this behavior
was attributed to the effect tension can have on
kinetochore-bound microtubules, with high tension
promoting polymerization (Maddox et al. 2003).
In the case of differentially sized microtubule bun-
dles attached to the kinetochore, the smaller bundle
will experience higher tension and therefore will
lengthen during anaphase, resulting in a shift of the
lagging chromosome away from the equator and
close to the pole connected to the thicker bundle
(Cimini et al. 2004) (Fig. 2B). In recent years, two
groups have highlighted a contribution of Aurora B
kinase, which localizes at the spindle midzone in
anaphase and can directly phosphorylate substrates
at the kinetochore of lagging chromosomes (Papini
et al. 2021), in minimizing the number of anaphase
lagging chromosomes resulting in chromosome
missegregation (Orr et al. 2021, Sen et al. 2021),
although there is still some debate on the exact
mechanism by which this happens [reviewed in
(Maiato and Silva 2023)]. An “anaphase correction
mechanism” was also invoked in an earlier study in
mouse oocytes showing a ten-fold ratio between the
number of lagging chromosomes in anaphase II and
the number of chromosomes that eventually misseg-
regated (Kouznetsova et al. 2019). Similar observa-
tions were also reported for anaphase II in insect
spermatocytes (Janicke et al. 2007). In at least one
cell line, it was shown that in cases in which a lag-
ging chromosome is incorporated into the main
nucleus, this is often the correct nucleus (Thomp-
son and Compton 2011).
Despite all the safety mechanisms described in
this section, some anaphase lagging chromosomes
will remain in proximity of the cell equator through
anaphase and beyond, and this can have dire conse-
quences [reviewed in (Krupina et al. 2021)], as dis-
cussed in the next section.
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Micronucleus formation and downstream effects
When an anaphase lagging chromosome persists in
proximity of the spindle equator through the end of
cell division, it invariably forms a micronucleus upon
mitotic exit (Cimini et al. 2002; Thompson and Comp-
ton 2011). It should be noted that anaphase lagging
chromosomes are not the only source of micronuclei,
which can also form from “polar chromosomes” that do
not align at the metaphase plate prior to anaphase onset
(Ferrandiz et al. 2022, Gomes et al. 2022) or fragments
of DNA resulting from pre-mitotic DNA damage or
chromosome bridges that break during cell division
(Hennig et al. 1988, Hoffelder et al. 2004, Pampalona
et al. 2016, Schuler et al. 1997, Umbreit et al. 2020).
Over the past decade, many studies have focused on
the biology of micronuclei and on the fate of cells that
enter mitosis with a micronucleus (Crasta et al. 2012,
Hatch et al. 2013, He et al. 2019, Liu et al. 2018, Soto
et al. 2018, Vazquez-Diez et al. 2016). Most of these
studies used experimental protocols that resulted in
increased rates of anaphase lagging chromosomes as a
way to increase the number of analyzable micronuclei
(Crasta et al. 2012, Hatch et al. 2013, He et al. 2019,
Liu et al. 2018, Soto et al. 2018). Thus, a wealth of
knowledge has been accumulated on the biology, fate,
and impact of lagging chromosome-derived micronu-
clei. An early study showed that micronuclei accumu-
lated DNA damage due to defective transport of DNA
repair enzymes and other proteins inside the micronu-
cleus (Crasta et al. 2012). Similar findings were also
reported in mouse embryos, where micronuclei dis-
played nuclear envelope defects, accumulation of DNA
damage, and impaired kinetochore assembly, which
in turn led to persistent missegregation of the micro-
nucleated chromosome (Vazquez-Diez et al. 2016).
These findings are consistent with earlier studies that
analyzed micronuclei containing chromosome frag-
ments instead of whole chromosomes. For instance,
micronuclei emerging from chromosome bridge break-
age were shown to display defective nuclear pore
complex assembly, nuclear import defects, reduced
transcriptional activity, and chromatin fragmentation
(Hoffelder et al. 2004). Similarly, radiation-induced
micronuclei (which likely contain DNA fragments)
displayed defective recruiting of DNA repair fac-
tors (Terradas et al. 2012, 2009), likely due to nuclear
envelope assembly defects arising from the inability
of lagging DNA to recruit non-core nuclear envelope
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Chromosome Res (2023) 31:18
proteins, including nuclear pore complexes, during
nuclear envelope reassembly upon mitotic exit (Liu
et al. 2018). Consequently, micronuclei fail to properly
import key proteins that are necessary for the integrity
of their nuclear envelope and DNA.
Micronuclei were also shown to frequently expe-
rience nuclear envelope rupture (Hatch et al. 2013;
Maciejowski and Hatch 2020). A recent study
showed that micronuclei containing larger and more
gene-dense chromosomes are less likely to rupture
(Mammel et al. 2022), but the chance of membrane
rupture is never zero. When the micronucleus enve-
lope ruptures, the micronuclear DNA can become
exposed to cytoplasmic content, including nucle-
ases that can digest and fragment the DNA, simi-
larly to what was observed for persistent chromatin
bridges (Maciejowski et al. 2015). Defective DNA
metabolism (e.g., defective DNA repair and repli-
cation) within micronuclei and micronucleus mem-
brane rupture have been proposed as the underlying
causes of the “chromosome pulverization” reported
in micronucleated cells that reenter mitosis (Crasta
et al. 2012; Kato and Sandberg 1968). Importantly,
this chromosome pulverization is believed to be a
precursor of chromothripsis, a catastrophic event in
which a single chromosome or chromosome region
undergoes shattering and re-stitching to gener-
ate a highly rearranged genomic region and that is
detected in a substantial number of tumors (Cortes-
Ciriano et al. 2020, Stephens et al. 2011). Several
studies have shown clear evidence of chromothripsis
being triggered by encapsulation of chromosomes
into micronuclei (Ly et al. 2019, 2017; Zhang et al.
2015). Ruptured micronuclei were also shown to
activate the cGAS-STING pathway responsible for
cellular immune response to cytosolic DNA (Hard-
ing et al. 2017; Maciejowski and Hatch 2020; Mac-
kenzie et al. 2017), and this was in turn shown to
promote cancer progression by driving metastasis
(Bakhoum et al. 2018; Krupina et al. 2021; Macie-
jowski and Hatch 2020).
Finally, even when they remain intact and trans-
port of DNA repair factors is not impaired, micronu-
clei were shown to display abnormal mitotic behavior
when the micronucleated cell reentered mitosis (He
et al. 2019; Soto et al. 2018). This was due to a num-
ber of mechanisms, including defective or delayed
chromosome condensation and defective recruitment
of kinetochore proteins (He et al. 2019; Soto et al.
Chromosome Res (2023) 31:18
2018). Due to defective chromosome condensation,
micronuclear DNA could also become trapped within
the cleavage furrow and cause it to regress (He et al.
2019), resulting in the formation of a binucleate,
tetraploid cell. This is relevant because tetraploidy
has been shown to promote tumorigenesis (Fujiwara
et al. 2005; Nguyen et al. 2009) and whole genome
doubling is a common event during tumor evolution
(Carter et al. 2012; Zack et al. 2013).
In conclusion, micronuclei arising from anaphase
lagging chromosomes represent a serious threat to
genomic stability. This is true even if the lagging
chromosome segregates to and forms a micronucleus
in the correct daughter cell.
Concluding remarks
The study that identified merotelic kinetochore
attachment as the underlying cause of anaphase lag-
ging chromosomes represented an important juncture.
The mitosis community recognized the importance of
aneuploidy and chromosomal instability (CIN) and
embarked on investigations aimed at determining the
contribution of anaphase lagging chromosomes to
CIN in cancer cells (Bakhoum et al. 2014; Thompson
and Compton 2008), examining what mechanisms
may enhance the formation of merotelic attachments
(Ganem et al. 2009; Kline-Smith et al. 2004; Silk-
worth et al. 2012; Silkworth et al. 2009) or promote
their correction (Bakhoum et al. 2009b; Cimini et al.
2006; DeLuca et al. 2006; Knowlton et al. 2006), and
explore the long-term consequences of lagging chro-
mosomes (Crasta et al. 2012; He et al. 2019; Ly et al.
2019; Ly et al. 2017; Soto et al. 2018; Vazquez-Diez
et al. 2016; Zhang et al. 2015).
What we have learned in recent years about the fate
of lagging chromosomes and the micronuclei derived
from them reinforces the traditional view that these
structures are good biomarkers of genotoxic dam-
age (Fenech et al. 2020), highlighting that this story
has come full circle. We have learned a tremendous
amount of detail along the way and this will serve the
scientific community well as we try to fill the remain-
ing gaps and answer new questions.
As described earlier, there is still substantial debate
on the exact mechanisms that contribute to correction
of kinetochore mis-attachments, including merotely,
and this area of investigation will likely continue to be
Page 9 of 15 18
prolific. A specific question that remains unanswered
is whether a correlation exists between chromosome
identity and efficiency of mis-attachment correction.
Such a bias could explain recent reports that certain
human chromosomes are more prone to missegregate
than others (Bochtler et al. 2019; Klaasen et al. 2022;
Worrall et al. 2018). It will be interesting to investi-
gate whether this bias in chromosome missegregation
depends on a bias in formation and/or correction of
merotelic attachments.
It will also be important to gain a deeper under-
standing of how merotelic kinetochore attachment
contributes to cancer. It is clear that lagging chromo-
somes are only one type of chromosome segregation
errors found in cancer cells. It will be important to
focus on examining tumor patient tissues and other
experimental systems to better understand the contri-
bution of different types of mitotic defects and their
inter-connections. For instance, chromosome bridges,
which are often seen in tumor cells (Bakhoum et al.
2011; Bolhaqueiro et al. 2019; Zaki et al. 2014), may
arise as a result of DNA damage in a micronucleus
that originally formed from a lagging chromosome.
This type of temporal information cannot be obtained
from analyzing fixed tissue samples and new strate-
gies may be needed to study the evolution of cell pop-
ulations in order to understand these interdependen-
cies. Such studies would untangle the many persistent
questions regarding the role of aneuploidy in cancer.
Indeed, although it is generally acknowledged that
aneuploidy is a cancer hallmark, the exact role of ane-
uploidy has not been fully elucidated and how ane-
uploidy accumulates in cancer cells is still unclear.
Notably, the recently described link between
micronuclei and activation of the cGAS-STING path-
way and the evidence that activation of this pathway
can promote tumorigenic phenotypes (Bakhoum
et al. 2018; Harding et al. 2017; Krupina et al. 2021;
Maciejowski and Hatch 2020; Mackenzie et al. 2017)
opens up new areas of investigation and opportunities
to exploit the phenotypic traits of CIN cancers and
their immune signature for therapeutic purposes.
Finally, there have been, over the years, reports
on a potential interplay between canonical tumor
suppressor genes or oncogenes, including P53, P21,
MYC, K-RAS, RAF/MEK/ERK, and fidelity of mitotic
chromosome segregation, tolerance to aneuploidy,
and CIN (Annibali et al. 2014; Giaretti 1997; Gron-
roos and Lopez-Garcia 2018; Kreis et al. 2014; Littler
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 18 Page 10 of 15
et al. 2019; Perera and Venkitaraman 2016; Rizzotto
et al. 2021; Zerbib et al. 2023). However, while there
is substantial evidence that P53 protects cells from
aneuploidy, tetraploidy, and CIN (Rizzotto et al.
2021), the relation between other tumor suppressor
genes or oncogenes and chromosome missegregation,
ploidy alterations, and CIN remains largely unex-
plored. Future studies in this area promise to uncover
new molecular details on the mechanisms of accurate
vs. defective chromosome segregation, bring to light
pathways to CIN and CIN tolerance, and possibly
inform new therapeutic strategies for cancer and other
diseases.
Acknowledgements I would like to acknowledge all mem-
bers of the Cimini lab for helpful discussions and careful read-
ing of the manuscript.
Authors’ contributions DC conceived the idea, performed
the literature search, wrote the manuscript, and prepared the
figures.
Funding Work in the Cimini lab is supported by grant
#R01GM140042 from the U.S. National Institutes of Health.
Additional funding is provided by a Dean’s Discovery Fund
award to DC from the Virginia Tech College of Science.
Data availability No original data or material are reported in
this review article.
Declarations
Ethical approval/consents This is a review article. As such,
no original research involving humans and/or animals is pre-
sented.
Competing interests The author has no competing interests
or other interests that might be perceived to influence the dis-
cussion reported in this paper.
References
Akiyoshi B, Sarangapani KK, Powers AF, Nelson CR, Reichow
SL, Arellano-Santoyo H, Gonen T, Ranish JA, Asbury
CL, Biggins S (2010) Tension directly stabilizes recon-
stituted kinetochore-microtubule attachments. Nature
468:576–579. https://​doi.​org/​10.​1038/​natur​e09594
Andrews PD, Ovechkina Y, Morrice N, Wagenbach M, Duncan
K, Wordeman L, Swedlow JR (2004) Aurora B regulates
MCAK at the mitotic centromere. Dev Cell 6:253–268.
https://​doi.​org/​10.​1016/​s1534-​5807(04)​00025-5
Annibali D, Whitfield JR, Favuzzi E, Jauset T, Serrano E, Cuar-
tas I, Redondo-Campos S, Folch G, Gonzalez-Junca A,
Vol:. (1234567890)
13
Chromosome Res (2023) 31:18
Sodir NM, Masso-Valles D, Beaulieu ME, Swigart LB,
Mc Gee MM, Somma MP, Nasi S, Seoane J, Evan GI,
Soucek L (2014) Myc inhibition is effective against gli-
oma and reveals a role for Myc in proficient mitosis. Nat
Commun 5:4632. https://​doi.​org/​10.​1038/​ncomm​s5632
Antoccia A, Degrassi F, Battistoni A, Ciliutti P, Tanzarella C
(1991) In vitro micronucleus test with kinetochore stain-
ing: evaluation of test performance. Mutagenesis 6:319–
324. https://​doi.​org/​10.​1093/​mutage/​6.4.​319
Bakhoum SF, Danilova OV, Kaur P, Levy NB, Compton DA
(2011) Chromosomal instability substantiates poor prog-
nosis in patients with diffuse large B-cell lymphoma.
Clin Cancer Res 17:7704–7711. https://​doi.​org/​10.​1158/​
1078-​0432.​CCR-​11-​2049
Bakhoum SF, Genovese G, Compton DA (2009a) Deviant
kinetochore microtubule dynamics underlie chromo-
somal instability. Curr Biol 19:1937–1942. https://​doi.​
org/​10.​1016/j.​cub.​2009.​09.​055
Bakhoum SF, Kabeche L, Wood MD, Laucius CD, Qu D,
Laughney AM, Reynolds GE, Louie RJ, Phillips J, Chan
DA, Zaki BI, Murnane JP, Petritsch C, Compton DA
(2015) Numerical chromosomal instability mediates sus-
ceptibility to radiation treatment. Nat Commun 6:5990.
https://​doi.​org/​10.​1038/​ncomm​s6990
Bakhoum SF, Ngo B, Laughney AM, Cavallo JA, Murphy CJ,
Ly P, Shah P, Sriram RK, Watkins TBK, Taunk NK,
Duran M, Pauli C, Shaw C, Chadalavada K, Rajasekhar
VK, Genovese G, Venkatesan S, Birkbak NJ, McGrana-
han N, Lundquist M, LaPlant Q, Healey JH, Elemento O,
Chung CH, Lee NY, Imielenski M, Nanjangud G, Pe’er
D, Cleveland DW, Powell SN, Lammerding J, Swanton
C, Cantley LC (2018) Chromosomal instability drives
metastasis through a cytosolic DNA response. Nature
553:467–472. https://​doi.​org/​10.​1038/​natur​e25432
Bakhoum SF, Silkworth WT, Nardi IK, Nicholson JM, Comp-
ton DA, Cimini D (2014) The mitotic origin of chromo-
somal instability. Curr Biol 24:R148-149. https://​doi.​org/​
10.​1016/j.​cub.​2014.​01.​019
Bakhoum SF, Thompson SL, Manning AL, Compton DA
(2009b) Genome stability is ensured by temporal con-
trol of kinetochore-microtubule dynamics. Nat Cell Biol
11:27–35. https://​doi.​org/​10.​1038/​ncb18​09
Baudoin NC, Cimini D (2018a) Chromosome Segregation:
The Bigger They Come, the Harder They Fall. Curr Biol
28:R665–R667. https://​doi.​org/​10.​1016/j.​cub.​2018.​04.​036
Baudoin NC, Cimini D (2018b) A guide to classifying mitotic
stages and mitotic defects in fixed cells. Chromosoma
127:215–227. https://​doi.​org/​10.​1007/​s00412-​018-​0660-2
Biggins S, Severin FF, Bhalla N, Sassoon I, Hyman AA, Mur-
ray AW (1999) The conserved protein kinase Ipl1 regu-
lates microtubule binding to kinetochores in budding
yeast. Genes Dev 13:532–544. https://​doi.​org/​10.​1101/​
gad.​13.5.​532
Bochtler T, Kartal-Kaess M, Granzow M, Hielscher T, Cosenza
MR, Herold-Mende C, Jauch A, Kramer A (2019) Micro-
nucleus formation in human cancer cells is biased by
chromosome size. Genes Chromosomes Cancer 58:392–
395. https://​doi.​org/​10.​1002/​gcc.​22707
Bolhaqueiro ACF, Ponsioen B, Bakker B, Klaasen SJ, Kucuk-
kose E, van Jaarsveld RH, Vivie J, Verlaan-Klink I, Hami
N, Spierings DCJ, Sasaki N, Dutta D, Boj SF, Vries RGJ,
Chromosome Res (2023) 31:18
Lansdorp PM, van de Wetering M, van Oudenaarden
A, Clevers H, Kranenburg O, Foijer F, Snippert HJG,
Kops G (2019) Ongoing chromosomal instability and
karyotype evolution in human colorectal cancer orga-
noids. Nat Genet 51:824–834. https://​doi.​org/​10.​1038/​
s41588-​019-​0399-6
Bonassi S, Coskun E, Ceppi M, Lando C, Bolognesi C, Burgaz
S, Holland N, Kirsh-Volders M, Knasmueller S, Zeiger
E, Carnesoltas D, Cavallo D, da Silva J, de Andrade VM,
Demircigil GC, Dominguez Odio A, Donmez-Altuntas H,
Gattas G, Giri A, Giri S, Gomez-Meda B, Gomez-Arroyo
S, Hadjidekova V, Haveric A, Kamboj M, Kurteshi K, Mar-
tino-Roth MG, Montero Montoya R, Nersesyan A, Pastor-
Benito S, Favero Salvadori DM, Shaposhnikova A, Stopper
H, Thomas P, Torres-Bugarin O, Yadav AS, Zuniga Gonza-
lez G, Fenech M (2011a) The HUman MicroNucleus pro-
ject on eXfoLiated buccal cells (HUMN(XL)): the role of
life-style, host factors, occupational exposures, health status,
and assay protocol. Mutat Res 728:88–97. https://​doi.​org/​
10.​1016/j.​mrrev.​2011.​06.​005
Bonassi S, El-Zein R, Bolognesi C, Fenech M (2011b) Micro-
nuclei frequency in peripheral blood lymphocytes and
cancer risk: evidence from human studies. Mutagenesis
26:93–100. https://​doi.​org/​10.​1093/​mutage/​geq075
Bonatti S, Cavalieri Z, Viaggi S, Abbondandolo A (1992) The
analysis of 10 potential spindle poisons for their ability
to induce CREST-positive micronuclei in human diploid
fibroblasts. Mutagenesis 7:111–114. https://​doi.​org/​10.​
1093/​mutage/​7.2.​111
Brinkley BR (2001) Managing the centrosome numbers game:
from chaos to stability in cancer cell division. Trends
Cell Biol 11:18–21. https://​doi.​org/​10.​1016/​s0962-​
8924(00)​01872-9
Broad AJ, DeLuca JG (2020) The right place at the right time:
Aurora B kinase localization to centromeres and kineto-
chores. Essays Biochem 64:299–311. https://​doi.​org/​10.​
1042/​EBC20​190081
Carter SL, Cibulskis K, Helman E, McKenna A, Shen H, Zack
T, Laird PW, Onofrio RC, Winckler W, Weir BA, Ber-
oukhim R, Pellman D, Levine DA, Lander ES, Meyer-
son M, Getz G (2012) Absolute quantification of somatic
DNA alterations in human cancer. Nat Biotechnol
30:413–421. https://​doi.​org/​10.​1038/​nbt.​2203
Catalan J, Falck GC, Norppa H (2000) The X chromosome
frequently lags behind in female lymphocyte anaphase.
Am J Hum Genet 66:687–691. https://​doi.​org/​10.​1086/​
302769
Cheeseman IM, Chappie JS, Wilson-Kubalek EM, Desai A
(2006) The conserved KMN network constitutes the
core microtubule-binding site of the kinetochore. Cell
127:983–997. https://​doi.​org/​10.​1016/j.​cell.​2006.​09.​039
Chen GY, Renda F, Zhang H, Gokden A, Wu DZ, Chenoweth
DM, Khodjakov A, Lampson MA (2021) Tension pro-
motes kinetochore-microtubule release by Aurora B
kinase. J Cell Biol 22010.1083/jcb.202007030
Cheng JM, Li J, Tang JX, Hao XX, Wang ZP, Sun TC, Wang
XX, Zhang Y, Chen SR, Liu YX (2017) Merotelic
kinetochore attachment in oocyte meiosis II causes
sister chromatids segregation errors in aged mice. Cell
Cycle 16:1404–1413. https://​doi.​org/​10.​1080/​15384​
101.​2017.​13274​88
Page 11 of 15 18
Ciferri C, Pasqualato S, Screpanti E, Varetti G, Santaguida
S, Dos Reis G, Maiolica A, Polka J, De Luca JG, De
Wulf P, Salek M, Rappsilber J, Moores CA, Salmon
ED, Musacchio A (2008) Implications for kinetochore-
microtubule attachment from the structure of an engi-
neered Ndc80 complex. Cell 133:427–439. https://​doi.​
org/​10.​1016/j.​cell.​2008.​03.​020
Cimini D, Antoccia A, Tanzarella C, Degrassi F (1997)
Topoisomerase II inhibition in mitosis produces
numerical and structural chromosomal aberrations in
human fibroblasts. Cytogenet Cell Genet 76:61–67.
https://​doi.​org/​10.​1159/​00013​4517
Cimini D, Cameron LA, Salmon ED (2004) Anaphase spin-
dle mechanics prevent mis-segregation of meroteli-
cally oriented chromosomes. Curr Biol 14:2149–2155.
https://​doi.​org/​10.​1016/j.​cub.​2004.​11.​029
Cimini D, Fioravanti D, Salmon ED, Degrassi F (2002)
Merotelic kinetochore orientation versus chromosome
mono-orientation in the origin of lagging chromo-
somes in human primary cells. J Cell Sci 115:507–515.
https://​doi.​org/​10.​1242/​jcs.​115.3.​507
Cimini D, Howell B, Maddox P, Khodjakov A, Degrassi F,
Salmon ED (2001) Merotelic kinetochore orientation is
a major mechanism of aneuploidy in mitotic mamma-
lian tissue cells. J Cell Biol 153:517–527. https://​doi.​
org/​10.​1083/​jcb.​153.3.​517
Cimini D, Moree B, Canman JC, Salmon ED (2003) Mero-
telic kinetochore orientation occurs frequently dur-
ing early mitosis in mammalian tissue cells and error
correction is achieved by two different mechanisms. J
Cell Sci 116:4213–4225. https://​doi.​org/​10.​1242/​jcs.​
00716
Cimini D, Tanzarella C, Degrassi F (1999) Differences in
malsegregation rates obtained by scoring ana-telo-
phases or binucleate cells. Mutagenesis 14:563–568.
https://​doi.​org/​10.​1093/​mutage/​14.6.​563
Cimini D, Wan X, Hirel CB, Salmon ED (2006) Aurora kinase
promotes turnover of kinetochore microtubules to reduce
chromosome segregation errors. Curr Biol 16:1711–
1718. https://​doi.​org/​10.​1016/j.​cub.​2006.​07.​022
Cortes-Ciriano I, Lee JJ, Xi R, Jain D, Jung YL, Yang L, Gor-
denin D, Klimczak LJ, Zhang CZ, Pellman DS, Group
PSVW, Park PJ, Consortium P (2020) Comprehensive
analysis of chromothripsis in 2,658 human cancers using
whole-genome sequencing. Nat Genet 52:331–341.
https://​doi.​org/​10.​1038/​s41588-​019-​0576-7
Crasta K, Ganem NJ, Dagher R, Lantermann AB, Ivanova
EV, Pan Y, Nezi L, Protopopov A, Chowdhury D, Pell-
man D (2012) DNA breaks and chromosome pulveriza-
tion from errors in mitosis. Nature 482:53–58. https://​
doi.​org/​10.​1038/​natur​e10802
de Regt AK, Clark CJ, Asbury CL, Biggins S (2022) Tension
can directly suppress Aurora B kinase-triggered release
of kinetochore-microtubule attachments. Nat Commun
13:2152. https://​doi.​org/​10.​1038/​s41467-​022-​29542-8
Degrassi F, Tanzarella C (1988) Immunofluorescent staining of
kinetochores in micronuclei: a new assay for the detec-
tion of aneuploidy. Mutat Res 203:339–345. https://​doi.​
org/​10.​1016/​0165-​1161(88)​90030-1
DeLuca JG, Gall WE, Ciferri C, Cimini D, Musacchio A,
Salmon ED (2006) Kinetochore microtubule dynamics
Vol.: (0123456789)
13
 18 Page 12 of 15
and attachment stability are regulated by Hec1. Cell
127:969–982. https://​doi.​org/​10.​1016/j.​cell.​2006.​09.​047
Deng C, Ya A, Compton DA, Godek KM (2023) A pluripo-
tent developmental state confers a low fidelity of chro-
mosome segregation. Stem Cell Reports 18:475–488.
https://​doi.​org/​10.​1016/j.​stemcr.​2022.​12.​008
Drpic D, Almeida AC, Aguiar P, Renda F, Damas J, Lewin
HA, Larkin DM, Khodjakov A, Maiato H (2018) Chro-
mosome Segregation Is Biased by Kinetochore Size.
Curr Biol 28:1344-1356 e1345. https://​doi.​org/​10.​1016/j.​
cub.​2018.​03.​023
Falck GC, Catalan J, Norppa H (2002) Nature of anaphase
laggards and micronuclei in female cytokinesis-blocked
lymphocytes. Mutagenesis 17:111–117. https://​doi.​org/​
10.​1093/​mutage/​17.2.​111
Fenech M (2000) The in vitro micronucleus technique. Mutat Res
455:81–95. https://​doi.​org/​10.​1016/​s0027-​5107(00)​00065-8
Fenech M, Knasmueller S, Bolognesi C, Holland N, Bonassi
S, Kirsch-Volders M (2020) Micronuclei as biomarkers
of DNA damage, aneuploidy, inducers of chromosomal
hypermutation and as sources of pro-inflammatory
DNA in humans. Mutat Res Rev Mutat Res 786:108342.
https://​doi.​org/​10.​1016/j.​mrrev.​2020.​108342
Fenech M, Morley AA (1989) Kinetochore detection in micro-
nuclei: an alternative method for measuring chromosome
loss. Mutagenesis 4:98–104. https://​doi.​org/​10.​1093/​
mutage/​4.2.​98
Ferrandiz N, Downie L, Starling GP, Royle SJ (2022) Endo-
membranes promote chromosome missegregation by
ensheathing misaligned chromosomes. J Cell Biol
22110.1083/jcb.202203021
Fujiwara T, Bandi M, Nitta M, Ivanova EV, Bronson RT,
Pellman D (2005) Cytokinesis failure generating tetra-
ploids promotes tumorigenesis in p53-null cells. Nature
437:1043–1047
Ganem NJ, Godinho SA, Pellman D (2009) A mechanism link-
ing extra centrosomes to chromosomal instability. Nature
460:278–282. https://​doi.​org/​10.​1038/​natur​e08136
Giaretti W (1997) Aneuploidy mechanisms in human colorec-
tal preneoplastic lesions and Barrett’s esophagus. Is there
a role for K-ras and p53 mutations? Anal Cell Pathol
15:99–117. https://​doi.​org/​10.​1155/​1997/​264135
Gisselsson D, Bjork J, Hoglund M, Mertens F, Dal Cin P, Aker-
man M, Mandahl N (2001) Abnormal nuclear shape in solid
tumors reflects mitotic instability. Am J Pathol 158:199–
206. https://​doi.​org/​10.​1016/​S0002-​9440(10)​63958-2
Gisselsson D, Palsson E, Yu C, Mertens F, Mandahl N (2004)
Mitotic instability associated with late genomic changes
in bone and soft tissue tumours. Cancer Lett 206:69–76.
https://​doi.​org/​10.​1016/j.​canlet.​2003.​10.​022
Gomes AM, Orr B, Novais-Cruz M, De Sousa F, Macario-Mon-
teiro J, Lemos C, Ferras C, Maiato H (2022) Micronuclei
from misaligned chromosomes that satisfy the spindle
assembly checkpoint in cancer cells. Curr Biol 32:4240-
4254 e4245. https://​doi.​org/​10.​1016/j.​cub.​2022.​08.​026
Gregan J, Polakova S, Zhang L, Tolic-Norrelykke IM, Cimini
D (2011) Merotelic kinetochore attachment: causes and
effects. Trends Cell Biol 21:374–381. https://​doi.​org/​10.​
1016/j.​tcb.​2011.​01.​003
Gregan J, Riedel CG, Pidoux AL, Katou Y, Rumpf C,
Schleiffer A, Kearsey SE, Shirahige K, Allshire RC,
Vol:. (1234567890)
13
Chromosome Res (2023) 31:18
Nasmyth K (2007) The kinetochore proteins Pcs1 and
Mde4 and heterochromatin are required to prevent mero-
telic orientation. Curr Biol 17:1190–1200. https://​doi.​
org/​10.​1016/j.​cub.​2007.​06.​044
Gronroos E, Lopez-Garcia C (2018) Tolerance of Chromo-
somal Instability in Cancer: Mechanisms and Therapeu-
tic Opportunities. Cancer Res 78:6529–6535. https://​doi.​
org/​10.​1158/​0008-​5472.​CAN-​18-​1958
Gustavino B, Degrassi F, Filipponi R, Modesti D, Tanzarella C,
Rizzoni M (1994) Mitotic indirect non-disjunction in phyto-
hemagglutinin stimulated human lymphocytes. Mutagenesis
9:17–21. https://​doi.​org/​10.​1093/​mutage/​9.1.​17
Harding SM, Benci JL, Irianto J, Discher DE, Minn AJ, Green-
berg RA (2017) Mitotic progression following DNA dam-
age enables pattern recognition within micronuclei. Nature
548:466–470. https://​doi.​org/​10.​1038/​natur​e23470
Hatch EM, Fischer AH, Deerinck TJ, Hetzer MW (2013) Cata-
strophic nuclear envelope collapse in cancer cell micronuclei.
Cell 154:47–60. https://​doi.​org/​10.​1016/j.​cell.​2013.​06.​007
He B, Gnawali N, Hinman AW, Mattingly AJ, Osimani A,
Cimini D (2019) Chromosomes missegregated into
micronuclei contribute to chromosomal instability by
missegregating at the next division. Oncotarget 10:2660–
2674. https://​doi.​org/​10.​18632/​oncot​arget.​26853
Hennig UG, Rudd NL, Hoar DI (1988) Kinetochore immu-
nofluorescence in micronuclei: a rapid method for the
in situ detection of aneuploidy and chromosome break-
age in human fibroblasts. Mutat Res 203:405–414.
https://​doi.​org/​10.​1016/​0165-​1161(88)​90013-1
Hoffelder DR, Luo L, Burke NA, Watkins SC, Gollin SM,
Saunders WS (2004) Resolution of anaphase bridges in
cancer cells. Chromosoma 112:389–397. https://​doi.​org/​
10.​1007/​s00412-​004-​0284-6
Hogstedt B, Karlsson A (1985) The size of micronuclei in
human lymphocytes varies according to inducing agent
used. Mutat Res 156:229–232. https://​doi.​org/​10.​1016/​
0165-​1218(85)​90067-9
Holubcova Z, Blayney M, Elder K, Schuh M (2015) Human
oocytes. Error-prone chromosome-mediated spindle
assembly favors chromosome segregation defects in
human oocytes. Science 348:1143–1147. https://​doi.​org/​
10.​1126/​scien​ce.​aaa95​29
Janicke MA, LaFountain JR Jr (1984) Malorientation in half-
bivalents at anaphase: analysis of autosomal laggards
in untreated, cold-treated, and cold-recovering crane fly
spermatocytes. J Cell Biol 98:859–869. https://​doi.​org/​
10.​1083/​jcb.​98.3.​859
Janicke MA, Lasko L, Oldenbourg R, LaFountain JR Jr (2007)
Chromosome malorientations after meiosis II arrest
cause nondisjunction. Mol Biol Cell 18:1645–1656.
https://​doi.​org/​10.​1091/​mbc.​e06-​10-​0963
Kaseda K, McAinsh AD, Cross RA (2012) Dual pathway spindle
assembly increases both the speed and the fidelity of mitosis.
Biol Open 1:12–18. https://​doi.​org/​10.​1242/​bio.​20110​12
Kato H, Sandberg AA (1968) Chromosome pulverization
in human cells with micronuclei. J Natl Cancer Inst
40:165–179
Khodjakov A, Cole RW, McEwen BF, Buttle KF, Rieder CL
(1997) Chromosome fragments possessing only one
kinetochore can congress to the spindle equator. J Cell
Biol 136:229–240. https://​doi.​org/​10.​1083/​jcb.​136.2.​229
Chromosome Res (2023) 31:18
Klaasen SJ, Truong MA, van Jaarsveld RH, Koprivec I, Sti-
mac V, de Vries SG, Risteski P, Kodba S, Vukusic K,
de Luca KL, Marques JF, Gerrits EM, Bakker B, Foijer
F, Kind J, Tolic IM, Lens SMA, Kops G (2022) Nuclear
chromosome locations dictate segregation error fre-
quencies. Nature 607:604–609. https://​doi.​org/​10.​1038/​
s41586-​022-​04938-0
Kline-Smith SL, Khodjakov A, Hergert P, Walczak CE (2004)
Depletion of centromeric MCAK leads to chromosome
congression and segregation defects due to improper
kinetochore attachments. Mol Biol Cell 15:1146–1159.
https://​doi.​org/​10.​1091/​mbc.​e03-​08-​0581
Knowlton AL, Lan W, Stukenberg PT (2006) Aurora B is
enriched at merotelic attachment sites, where it regulates
MCAK. Curr Biol 16:1705–1710. https://​doi.​org/​10.​
1016/j.​cub.​2006.​07.​057
Kouznetsova A, Hernandez-Hernandez A, Hoog C (2014)
Merotelic attachments allow alignment and stabiliza-
tion of chromatids in meiosis II oocytes. Nat Commun
5:4409. https://​doi.​org/​10.​1038/​ncomm​s5409
Kouznetsova A, Kitajima TS, Brismar H, Hoog C (2019) Post-
metaphase correction of aberrant kinetochore-micro-
tubule attachments in mammalian eggs. EMBO Rep
20:e47905. https://​doi.​org/​10.​15252/​embr.​20194​7905
Kreis NN, Sanhaji M, Rieger MA, Louwen F, Yuan J (2014)
p21Waf1/Cip1 deficiency causes multiple mitotic defects
in tumor cells. Oncogene 33:5716–5728. https://​doi.​org/​
10.​1038/​onc.​2013.​518
Krupina K, Goginashvili A, Cleveland DW (2021) Causes
and consequences of micronuclei. Curr Opin Cell Biol
70:91–99. https://​doi.​org/​10.​1016/j.​ceb.​2021.​01.​004
Ladrach KS, LaFountain JR Jr (1986) Malorientation and
abnormal segregation of chromosomes during recovery
from colcemid and nocodazole. Cell Motil Cytoskeleton
6:419–427. https://​doi.​org/​10.​1002/​cm.​97006​0407
Lan W, Zhang X, Kline-Smith SL, Rosasco SE, Barrett-Wilt
GA, Shabanowitz J, Hunt DF, Walczak CE, Stukenberg
PT (2004) Aurora B phosphorylates centromeric MCAK
and regulates its localization and microtubule depolym-
erization activity. Curr Biol 14:273–286. https://​doi.​org/​
10.​1016/j.​cub.​2004.​01.​055
Lara-Gonzalez P, Pines J, Desai A (2021) Spindle assem-
bly checkpoint activation and silencing at kinetochores.
Semin Cell Dev Biol 117:86–98. https://​doi.​org/​10.​
1016/j.​semcdb.​2021.​06.​009
Lengauer C, Kinzler KW, Vogelstein B (1997) Genetic insta-
bility in colorectal cancers. Nature 386:623–627. https://​
doi.​org/​10.​1038/​38662​3a0
Littler S, Sloss O, Geary B, Pierce A, Whetton AD, Taylor SS
(2019) Oncogenic MYC amplifies mitotic perturbations.
Open Biol 9:190136. https://​doi.​org/​10.​1098/​rsob.​190136
Liu S, Kwon M, Mannino M, Yang N, Renda F, Khodjakov A,
Pellman D (2018) Nuclear envelope assembly defects
link mitotic errors to chromothripsis. Nature 561:551–
555. https://​doi.​org/​10.​1038/​s41586-​018-​0534-z
Ly P, Brunner SF, Shoshani O, Kim DH, Lan W, Pyntik-
ova T, Flanagan AM, Behjati S, Page DC, Campbell
PJ, Cleveland DW (2019) Chromosome segregation
errors generate a diverse spectrum of simple and com-
plex genomic rearrangements. Nat Genet 51:705–715.
https://​doi.​org/​10.​1038/​s41588-​019-​0360-8
Page 13 of 15 18
Ly P, Teitz LS, Kim DH, Shoshani O, Skaletsky H, Fachi-
netti D, Page DC, Cleveland DW (2017) Selective Y
centromere inactivation triggers chromosome shatter-
ing in micronuclei and repair by non-homologous end
joining. Nat Cell Biol 19:68–75. https://​doi.​org/​10.​
1038/​ncb34​50
Maciejowski J, Hatch EM (2020) Nuclear Membrane Rup-
ture and Its Consequences. Annu Rev Cell Dev Biol
36:85–114. https://​doi.​org/​10.​1146/​annur​ev-​cellb​
io-​020520-​120627
Maciejowski J, Li Y, Bosco N, Campbell PJ, de Lange T (2015)
Chromothripsis and Kataegis Induced by Telomere Cri-
sis. Cell 163:1641–1654. https://​doi.​org/​10.​1016/j.​cell.​
2015.​11.​054
Mackenzie KJ, Carroll P, Martin CA, Murina O, Fluteau A,
Simpson DJ, Olova N, Sutcliffe H, Rainger JK, Leitch A,
Osborn RT, Wheeler AP, Nowotny M, Gilbert N, Chan-
dra T, Reijns MAM, Jackson AP (2017) cGAS surveil-
lance of micronuclei links genome instability to innate
immunity. Nature 548:461–465. https://​doi.​org/​10.​1038/​
natur​e23449
Maddox P, Straight A, Coughlin P, Mitchison TJ, Salmon ED
(2003) Direct observation of microtubule dynamics at
kinetochores in Xenopus extract spindles: implications
for spindle mechanics. J Cell Biol 162:377–382. https://​
doi.​org/​10.​1083/​jcb.​20030​1088
Maiato H, Silva S (2023) Double-checking chromosome segre-
gation. J Cell Biol 22210.1083/jcb.202301106
Mammel AE, Huang HZ, Gunn AL, Choo E, Hatch EM
(2022) Chromosome length and gene density contribute
to micronuclear membrane stability. Life Sci Alliance
510.26508/lsa.202101210
Maney T, Hunter AW, Wagenbach M, Wordeman L (1998)
Mitotic centromere-associated kinesin is important for
anaphase chromosome segregation. J Cell Biol 142:787–
801. https://​doi.​org/​10.​1083/​jcb.​142.3.​787
Mihajlovic AI, Haverfield J, FitzHarris G (2021) Distinct
classes of lagging chromosome underpin age-related
oocyte aneuploidy in mouse. Dev Cell 56:2273-2283
e2273. https://​doi.​org/​10.​1016/j.​devcel.​2021.​07.​022
Moroi Y, Peebles C, Fritzler MJ, Steigerwald J, Tan EM (1980)
Autoantibody to centromere (kinetochore) in sclero-
derma sera. Proc Natl Acad Sci U S A 77:1627–1631.
https://​doi.​org/​10.​1073/​pnas.​77.3.​1627
Nam HJ, van Deursen JM (2014) Cyclin B2 and p53 control
proper timing of centrosome separation. Nat Cell Biol
16:538–549. https://​doi.​org/​10.​1038/​ncb29​52
Nguyen HG, Makitalo M, Yang D, Chinnappan D, St Hilaire
C, Ravid K (2009) Deregulated Aurora-B induced tetra-
ploidy promotes tumorigenesis. FASEB J 23:2741–2748.
https://​doi.​org/​10.​1096/​fj.​09-​130963
Nicholson JM, Cimini D (2013) Cancer karyotypes: survival
of the fittest. Front Oncol 3:148. https://​doi.​org/​10.​3389/​
fonc.​2013.​00148
Orr B, De Sousa F, Gomes AM, Afonso O, Ferreira LT,
Figueiredo AC, Maiato H (2021) An anaphase surveil-
lance mechanism prevents micronuclei formation from
frequent chromosome segregation errors. Cell Rep
37:109783. https://​doi.​org/​10.​1016/j.​celrep.​2021.​109783
Orr B, Talje L, Liu Z, Kwok BH, Compton DA (2016)
Adaptive Resistance to an Inhibitor of Chromosomal
Vol.: (0123456789)
13
 18 Page 14 of 15
Instability in Human Cancer Cells. Cell Rep 17:1755–
1763. https://​doi.​org/​10.​1016/j.​celrep.​2016.​10.​030
Pampalona J, Roscioli E, Silkworth WT, Bowden B, Genesca A,
Tusell L, Cimini D (2016) Chromosome Bridges Main-
tain Kinetochore-Microtubule Attachment throughout
Mitosis and Rarely Break during Anaphase. PLoS One
11:e0147420. https://​doi.​org/​10.​1371/​journ​al.​pone.​01474​20
Papini D, Levasseur MD, Higgins JMG (2021) The Aurora B gra-
dient sustains kinetochore stability in anaphase. Cell Rep
37:109818. https://​doi.​org/​10.​1016/j.​celrep.​2021.​109818
Paul R, Wollman R, Silkworth WT, Nardi IK, Cimini D,
Mogilner A (2009) Computer simulations predict that
chromosome movements and rotations accelerate mitotic
spindle assembly without compromising accuracy. Proc
Natl Acad Sci U S A 106:15708–15713. https://​doi.​org/​
10.​1073/​pnas.​09082​61106
Perera D, Venkitaraman AR (2016) Oncogenic KRAS triggers
MAPK-dependent errors in mitosis and MYC-depend-
ent sensitivity to anti-mitotic agents. Sci Rep 6:29741.
https://​doi.​org/​10.​1038/​srep2​9741
Pidoux AL, Uzawa S, Perry PE, Cande WZ, Allshire RC
(2000) Live analysis of lagging chromosomes during
anaphase and their effect on spindle elongation rate in
fission yeast. J Cell Sci 113(Pt 23):4177–4191. https://​
doi.​org/​10.​1242/​jcs.​113.​23.​4177
Pincu M, Callisen H, Norman A (1985) DNA content of micro-
nuclei in human lymphocytes. Int J Radiat Biol Relat
Stud Phys Chem Med 47:423–432
Rajagopalan H, Lengauer C (2004) Aneuploidy and cancer.
Nature 432:338–341. https://​doi.​org/​10.​1038/​natur​e03099
Rieder CL, Cole RW, Khodjakov A, Sluder G (1995) The
checkpoint delaying anaphase in response to chromo-
some monoorientation is mediated by an inhibitory sig-
nal produced by unattached kinetochores. J Cell Biol
130:941–948. https://​doi.​org/​10.​1083/​jcb.​130.4.​941
Rieder CL, Schultz A, Cole R, Sluder G (1994) Anaphase
onset in vertebrate somatic cells is controlled by a check-
point that monitors sister kinetochore attachment to the
spindle. J Cell Biol 127:1301–1310. https://​doi.​org/​10.​
1083/​jcb.​127.5.​1301
Rizzoni M, Tanzarella C, Gustavino B, Degrassi F, Guarino
A, Vitagliano E (1989) Indirect mitotic nondisjunction
in Vicia faba and Chinese hamster cells. Chromosoma
97:339–346. https://​doi.​org/​10.​1007/​BF003​71976
Rizzotto D, Englmaier L, Villunger A (2021) At a Cross-
roads to Cancer: How p53-Induced Cell Fate Decisions
Secure Genome Integrity. Int J Mol Sci 2210.3390/
ijms221910883
Rumpf C, Cipak L, Schleiffer A, Pidoux A, Mechtler K, Tolic-
Norrelykke IM, Gregan J (2010) Laser microsurgery
provides evidence for merotelic kinetochore attachments
in fission yeast cells lacking Pcs1 or Clr4. Cell Cycle
9:3997–4004. https://​doi.​org/​10.​4161/​cc.9.​19.​13233
Sarangapani KK, Asbury CL (2014) Catch and release: how do
kinetochores hook the right microtubules during mitosis?
Trends Genet 30:150–159. https://​doi.​org/​10.​1016/j.​tig.​
2014.​02.​004
Schuler M, Rupa DS, Eastmond DA (1997) A critical evalu-
ation of centromeric labeling to distinguish micronuclei
induced by chromosomal loss and breakage in vitro.
Vol:. (1234567890)
13
Chromosome Res (2023) 31:18
Mutat Res 392:81–95. https://​doi.​org/​10.​1016/​s0165-​
1218(97)​00047-5
Sen O, Harrison JU, Burroughs NJ, McAinsh AD (2021) Kine-
tochore life histories reveal an Aurora-B-dependent error
correction mechanism in anaphase. Dev Cell 56:3082-
3099 e3085. https://​doi.​org/​10.​1016/j.​devcel.​2021.​10.​007
Shomper M, Lappa C, FitzHarris G (2014) Kinetochore microtu-
bule establishment is defective in oocytes from aged mice.
Cell Cycle 13:1171–1179. https://​doi.​org/​10.​4161/​cc.​28046
Silkworth WT, Cimini D (2012) Transient defects of mitotic
spindle geometry and chromosome segregation errors.
Cell Div 7:19. https://​doi.​org/​10.​1186/​1747-​1028-7-​19
Silkworth WT, Nardi IK, Paul R, Mogilner A, Cimini D (2012)
Timing of centrosome separation is important for accu-
rate chromosome segregation. Mol Biol Cell 23:401–
411. https://​doi.​org/​10.​1091/​mbc.​E11-​02-​0095
Silkworth WT, Nardi IK, Scholl LM, Cimini D (2009)
Multipolar spindle pole coalescence is a major source of
kinetochore mis-attachment and chromosome mis-segre-
gation in cancer cells. PLoS One 4:e6564. https://​doi.​org/​
10.​1371/​journ​al.​pone.​00065​64
Sluder G, Thompson EA, Miller FJ, Hayes J, Rieder CL (1997)
The checkpoint control for anaphase onset does not mon-
itor excess numbers of spindle poles or bipolar spindle
symmetry. J Cell Sci 110(Pt 4):421–429. https://​doi.​org/​
10.​1242/​jcs.​110.4.​421
Soto M, Garcia-Santisteban I, Krenning L, Medema RH,
Raaijmakers JA (2018) Chromosomes trapped in
micronuclei are liable to segregation errors. J Cell Sci
13110.1242/jcs.214742
Stear JH, Roth MB (2002) Characterization of HCP-6, a C.
elegans protein required to prevent chromosome twist-
ing and merotelic attachment. Genes Dev 16:1498–1508.
https://​doi.​org/​10.​1101/​gad.​989102
Stephens PJ, Greenman CD, Fu B, Yang F, Bignell GR, Mudie
LJ, Pleasance ED, Lau KW, Beare D, Stebbings LA,
McLaren S, Lin ML, McBride DJ, Varela I, Nik-Zainal S,
Leroy C, Jia M, Menzies A, Butler AP, Teague JW, Quail
MA, Burton J, Swerdlow H, Carter NP, Morsberger LA,
Iacobuzio-Donahue C, Follows GA, Green AR, Flanagan
AM, Stratton MR, Futreal PA, Campbell PJ (2011) Mas-
sive genomic rearrangement acquired in a single cata-
strophic event during cancer development. Cell 144:27–
40. https://​doi.​org/​10.​1016/j.​cell.​2010.​11.​055
Stewenius Y, Jin Y, Ora I, de Kraker J, Bras J, Frigyesi A, Alumets
J, Sandstedt B, Meeker AK, Gisselsson D (2007) Defec-
tive chromosome segregation and telomere dysfunction in
aggressive Wilms’ tumors. Clin Cancer Res 13:6593–6602.
https://​doi.​org/​10.​1158/​1078-​0432.​CCR-​07-​1081
Streffer C, van Beuningen D, Rebmann A (1985) DNA meas-
urements and micronuclei in human rectal carcinoma.
Radiat Med 3:147–150
Tanaka TU, Rachidi N, Janke C, Pereira G, Galova M, Schiebel
E, Stark MJ, Nasmyth K (2002) Evidence that the Ipl1-
Sli15 (Aurora kinase-INCENP) complex promotes chro-
mosome bi-orientation by altering kinetochore-spindle
pole connections. Cell 108:317–329. https://​doi.​org/​10.​
1016/​s0092-​8674(02)​00633-5
Terradas M, Martin M, Hernandez L, Tusell L, Genesca A
(2012) Nuclear envelope defects impede a proper response
Chromosome Res (2023) 31:18
to micronuclear DNA lesions. Mutat Res 729:35–40.
https://​doi.​org/​10.​1016/j.​mrfmmm.​2011.​09.​003
Terradas M, Martin M, Tusell L, Genesca A (2009) DNA
lesions sequestered in micronuclei induce a local defec-
tive-damage response. DNA Repair (amst) 8:1225–1234.
https://​doi.​org/​10.​1016/j.​dnarep.​2009.​07.​004
Thompson SL, Compton DA (2008) Examining the link
between chromosomal instability and aneuploidy in
human cells. J Cell Biol 180:665–672. https://​doi.​org/​10.​
1083/​jcb.​20071​2029
Thompson SL, Compton DA (2011) Chromosome missegrega-
tion in human cells arises through specific types of kine-
tochore-microtubule attachment errors. Proc Natl Acad
Sci U S A 108:17974–17978. https://​doi.​org/​10.​1073/​
pnas.​11097​20108
Thomson EJ, Perry PE (1988) The identification of micronucleated
chromosomes: a possible assay for aneuploidy. Mutagenesis
3:415–418. https://​doi.​org/​10.​1093/​mutage/​3.5.​415
Trivedi P, Zaytsev AV, Godzi M, Ataullakhanov FI, Grish-
chuk EL, Stukenberg PT (2019) The binding of
Borealin to microtubules underlies a tension inde-
pendent kinetochore-microtubule error correction path-
way. Nat Commun 10:682. https://​doi.​org/​10.​1038/​
s41467-​019-​08418-4
Tucker JB, Bonema SC, Garcia-Varela R, Denu RA, Hu Y,
McGregor SM, Burkard ME, Weaver BA (2023) Mis-
aligned Chromosomes are a Major Source of Chromo-
somal Instability in Breast Cancer. Cancer Res Commun
3:54–65. https://​doi.​org/​10.​1158/​2767-​9764.​CRC-​22-​0302
Umbreit NT, Zhang CZ, Lynch LD, Blaine LJ, Cheng AM,
Tourdot R, Sun L, Almubarak HF, Judge K, Mitchell TJ,
Spektor A, Pellman D (2020) Mechanisms generating can-
cer genome complexity from a single cell division error.
Science 368. https://​doi.​org/​10.​1126/​scien​ce.​aba07​12
Vazquez-Diez C, Yamagata K, Trivedi S, Haverfield J, FitzHar-
ris G (2016) Micronucleus formation causes perpetual
unilateral chromosome inheritance in mouse embryos.
Proc Natl Acad Sci U S A 113:626–631. https://​doi.​org/​
10.​1073/​pnas.​15176​28112
Venkatesan S, Angelova M, Puttick C, Zhai H, Caswell DR,
Lu WT, Dietzen M, Galanos P, Evangelou K, Bellelli R,
Lim EL, Watkins TBK, Rowan A, Teixeira VH, Zhao Y,
Chen H, Ngo B, Zalmas LP, Al Bakir M, Hobor S, Gron-
roos E, Pennycuick A, Nigro E, Campbell BB, Brown
WL, Akarca AU, Marafioti T, Wu MY, Howell M, Boul-
ton SJ, Bertoli C, Fenton TR, de Bruin RAM, Maya-
Mendoza A, Santoni-Rugiu E, Hynds RE, Gorgoulis
VG, Jamal-Hanjani M, McGranahan N, Harris RS, Janes
SM, Bartkova J, Bakhoum SF, Bartek J, Kanu N, Swan-
ton C, Consortium TR (2021) Induction of APOBEC3
Exacerbates DNA Replication Stress and Chromosomal
Instability in Early Breast and Lung Cancer Evolution.
Cancer Discov 11:2456–2473. https://​doi.​org/​10.​1158/​
2159-​8290.​CD-​20-​0725
Weaver BA, Cleveland DW (2006) Does aneuploidy cause can-
cer? Curr Opin Cell Biol 18:658–667. https://​doi.​org/​10.​
1016/j.​ceb.​2006.​10.​002
Wise DA, Brinkley BR (1997) Mitosis in cells with unreplicated
genomes (MUGs): spindle assembly and behavior of cen-
tromere fragments. Cell Motil Cytoskeleton 36:291–302.
Page 15 of 15 18
https://​doi.​org/​10.​1002/​(SICI)​1097-​0169(1997)​36:3%​
3c291::​AID-​CM9%​3e3.0.​CO;2-A
Worrall JT, Tamura N, Mazzagatti A, Shaikh N, van Lingen
T, Bakker B, Spierings DCJ, Vladimirou E, Foijer F,
McClelland SE (2018) Non-random Mis-segregation of
Human Chromosomes. Cell Rep 23:3366–3380. https://​
doi.​org/​10.​1016/j.​celrep.​2018.​05.​047
Yamamoto KI, Kikuchi Y (1980) A comparison of diameters
of micronuclei induced by clastogens and by spindle poi-
sons. Mutat Res 71:127–131. https://​doi.​org/​10.​1016/​
0027-​5107(80)​90012-3
Yu HG, Dawe RK (2000) Functional redundancy in the maize
meiotic kinetochore. J Cell Biol 151:131–142. https://​
doi.​org/​10.​1083/​jcb.​151.1.​131
Zack TI, Schumacher SE, Carter SL, Cherniack AD, Saksena G,
Tabak B, Lawrence MS, Zhsng CZ, Wala J, Mermel CH,
Sougnez C, Gabriel SB, Hernandez B, Shen H, Laird PW,
Getz G, Meyerson M, Beroukhim R (2013) Pan-cancer
patterns of somatic copy number alteration. Nat Genet
45:1134–1140. https://​doi.​org/​10.​1038/​ng.​2760
Zaki BI, Suriawinata AA, Eastman AR, Garner KM, Bakhoum
SF (2014) Chromosomal instability portends superior
response of rectal adenocarcinoma to chemoradiation
therapy. Cancer 120:1733–1742. https://​doi.​org/​10.​1002/​
cncr.​28656
Zerbib J, Ippolito MR, Eliezer Y, De Feudis G, Reuveni E,
Kadmon AS, Martin S, Viganò S, Leor G, Berstler J,
Laue K, Cohen-Sharir Y, Scorzoni S, Vazquez F, Ben-
David U, Santaguida S (2023) Human aneuploid cells
depend on the RAF/MEK/ERK pathway for overcoming
increased DNA damage. bioRxiv
Zhai Y, Kronebusch PJ, Borisy GG (1995) Kinetochore microtubule
dynamics and the metaphase-anaphase transition. J Cell Biol
131:721–734. https://​doi.​org/​10.​1083/​jcb.​131.3.​721
Zhang CZ, Spektor A, Cornils H, Francis JM, Jackson EK,
Liu S, Meyerson M, Pellman D (2015) Chromothripsis
from DNA damage in micronuclei. Nature 522:179–184.
https://​doi.​org/​10.​1038/​natur​e14493
Zhang X, Lan W, Ems-McClung SC, Stukenberg PT, Walczak
CE (2007) Aurora B phosphorylates multiple sites on
mitotic centromere-associated kinesin to spatially and
temporally regulate its function. Mol Biol Cell 18:3264–
3276. https://​doi.​org/​10.​1091/​mbc.​e07-​01-​0086
Zielinska AP, Holubcova Z, Blayney M, Elder K, Schuh M
(2015) Sister kinetochore splitting and precocious disinte-
gration of bivalents could explain the maternal age effect.
Elife 4:e11389. https://​doi.​org/​10.​7554/​eLife.​11389
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13