REVIEWS

Mitonuclear communication in
homeostasis and stress
Pedro M. Quirós*, Adrienne Mottis* and Johan Auwerx
Abstract | Mitochondria participate in crucial cellular processes such as energy harvesting and
intermediate metabolism. Although mitochondria possess their own genome — a vestige of their
bacterial origins and endosymbiotic evolution — most mitochondrial proteins are encoded in the
nucleus. The expression of the mitochondrial proteome hence requires tight coordination
between the two genomes to adapt mitochondrial function to the ever-changing cellular milieu.
In this Review, we focus on the pathways that coordinate the communication between
mitochondria and the nucleus during homeostasis and mitochondrial stress. These pathways
include nucleus-to‑mitochondria (anterograde) and mitochondria-to‑nucleus (retrograde)
communication, mitonuclear feedback signalling and proteostasis regulation, the integrated
stress response and non-cell-autonomous communication. We discuss how mitonuclear
communication safeguards cellular and organismal fitness and regulates lifespan.

Endosymbionts
Organisms that live within
another organism in a
symbiotic relationship.
Electron transport chain
(ETC). A group of protein
complexes that pass electrons
from one to another via redox
reactions coupled with the
transfer of protons across the
inner mitochondrial
membrane, creating a proton
gradient that drives
ATP synthesis.
Intermediate metabolism
The intermediate steps by
which nutrient molecules or
foodstuffs are metabolized
intracellularly.
Laboratory for Integrative
and Systems Physiology,
Ecole Polytechnique Fédérale
de Lausanne (EPFL),
CH‑1015, Lausanne,
Switzerland.
*These authors contributed
equally to this article.
Correspondence to J.A.
admin.auwerx@epfl.ch
doi:10.1038/nrm.2016.23
Published online 9 March 2016
Mitochondria are derived from α‑proteobacteria that
were engulfed by the precursor of modern eukaryotic cells
before evolving as endosymbionts over millions of years.
These organelles have maintained some of their ances‑
tral bacterial characteristics, such as a circular genome
and the capacity to produce ATP — mito­chondria are
the core of energy metabolism within the cell1. During
evolution, however, many of the proteo­bacterial genes
were pro­gressively transferred to the nuclear genome,
and mitochondria acquired new components and func‑
tions from the host cell, resulting in profound changes in
both the mitochondrial and the nuclear genome and pro‑
teome2. Out of more than 1,200 proteins that are present
in mitochondria, only 13 are encoded by the mammalian
mitochondrial DNA (mtDNA) — 12 in Caenorhabditis
elegans3,4. These 13 proteins constitute important compo‑
nents of all complexes of the electron transport chain (ETC)
except for complex II, the components of which are exclu‑
sively encoded within the nuclear genome. Therefore,
the nucleus and mito­chondria must continuously
­coordinate the transcription and translation, as well as
the t­ ranslocation and import, of ­mitochondrial proteins3.
Mitochondria are not only at the heart of cellular
energy harvesting, however, they also regulate many
aspects of intermediate metabolism, calcium buffering and
processes such as apoptosis. Consequently, mitochon‑
drial function is under tight nuclear control, through
so‑called ‘anterograde regulation’, which can decrease
or increase mitochondrial activity, as well as promoting
mito­chondrial biogenesis, depending on cellular needs.
Conversely, mitochondria can generate a ‘retrograde
response’ that signals to the nucleus to alter the expression
of nuclear genes to modify cellular function and repro‑
gramme cell metabolism. The integration of these antero­
grade (from nucleus to mitochondria) and retrograde
(from mitochondria to nucleus) signals — also known
as mitonuclear communication — constitutes a robust
network that helps cells to maintain homeostasis under
basal conditions and enables their adaptation to various
stressors. However, mitonuclear communication is most
often bidirectional and of a hormetic nature, combining
anterograde and retrograde signals. Thus, retrograde sig‑
nals that are generated in reaction to some mitochondrial
stressors trigger particular nuclear responses, which will
induce the speci­fic expression of certain mitochondrial
proteins; in turn, these proteins will resolve the original
perturbations in the mitochondria through ‘mitonuclear
feedback’. Depending on the severity and nature of the
initial stress signal, mitonuclear communication can
result in an ‘integrated stress response’ (ISR). The ISR
can also be induced by other stress signals, such as those
originating from endoplasmic reticulum (ER) stress,
triggering a global cellular response by decreasing pro‑
tein synthesis. Moreover, mitonuclear stress can also trig‑
ger a non-cell-autonomous response, which modulates
the function of distant cells to facilitate an organismal
response or adaptation to stress (FIG. 1).
Interest in understanding these complex mito­nuclear
responses has increased during the past decade, mainly
because of their key involvement in health and lifespan.
In this Review, we discuss the functional interaction
between mitochondria and the nucleus, with a particular

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Anterograde Retrograde mitochondrial transcription and translation are encoded

regulation response in the nucleus, and many of them are regulated by NRFs.
Mitochondrial transcription is controlled by mito‑
Nucleus
chondrial transcription and termination factors that
collabor­ate with the mitochondrial RNA polymerase to
Mitochondrion generate two polycistronic mitochondrial RNAs9, which
are translated by mitochondrial ribosomes in a process
Extracellular that is tightly regulated by nuclear-encoded factors and
communication Mitonuclear
feedback integrated into cellular processes10.
ER Certain nuclear receptors can also activate the expres‑
Integrated
sion of nuclear-encoded mitochondrial proteins. For
stress response example, some peroxisome proliferator-activated recep‑
tors (PPARs), such as PPARδ, stimulate the expression
Figure 1 | Mitonuclear communication. Mitochondria and
Nature Reviews | Molecular Cell
the nucleus communicate closely with each other. From a Biology of enzymes that are involved in mitochondrial fatty acid
‘mitocentric’ point of view, as illustrated here, signals sent oxidation, especially in the heart and muscle, in which this
from the nucleus to mitochondria constitute anterograde process is key for generating ATP11,12. Other PPARs have
regulation, whereas those sent by mitochondria to the similar functions in other tissues (reviewed in REF. 13).
nucleus are defined as the retrograde response. Although The oestrogen-related receptors (ERRs) ERRα, β and γ are
mitonuclear feedback responses can be orchestrated associated with the expression of nuclear-encoded mito‑
bidirectionally, they usually originate from mitochondria chondrial proteins that are involved in the tricarboxylic
and consequently induce a nuclear response that is acid (TCA) cycle, OXPHOS and fatty acid oxidation14,15.
specifically geared towards the mitochondria. The Fine-tuning of the action of these transcription fac‑
integrated stress response is a general cellular stress
tors is coordinated by co-activators or co-­repressors16.
pathway that controls cytosolic protein synthesis; this stress
response can be triggered in mitochondria, in the
Within the context of mitochondria, PPARγ c­ o‑
endoplasmic reticulum (ER) and in the cytosol. Mitonuclear activator 1α (PGC1α), PGC1β and the PGC-related
communication can also be extracellular, as mitochondria co-­activator PRC are known to activate the NRF, ERR
can send extracellular cues (known as mitokines), which and PPAR factors to induce mitochondrial bio­genesis17,18,
affect nuclear regulation in a non-cell-autonomous manner. whereas ­co-repressors such as nuclear receptor co-­
repressor 1 or r­ eceptor-interacting protein 140 can inhibit
their function19,20.
emphasis on how this interaction is affected by cellu‑ These anterograde signals are activated by upstream
lar stress. We also describe the different pathways that sensors that detect changes in metabolic conditions.
coordi­nate mitonuclear communication and how this A decrease in ATP synthesis, as seen during exercise and
process influences cellular function and organismal caloric restriction, increases the AMP/ATP ratio and acti‑
homeostasis and lifespan. vates AMP-activated protein kinase (AMPK), which, in
turn, increases cellular NAD+ levels. Increased NAD+
Anterograde regulation levels lead to activation of sirtuin 1 (SIRT1)21,22 — a
Mitochondrial function is controlled by the nucleus NAD+-dependent deacetylase that positively regulates
through anterograde signals, which allows these PGC1α — and subsequent activation of mitochondrial
organelles to adapt to the cellular milieu. This regu‑ energy metabolism and biogenesis23. Increases in levels
lation is based mainly on the expression of nuclear-­ of calcium ions (Ca2+) during exercise activate PGC1α
encoded mitochondrial proteins that induce mtDNA — either directly through Ca2+/calmodulin-­dependent
gene expression, and on several transcription factors protein kinase type IV (CaMKIV)24 or through the
and co‑­regulators that regulate the expression of the activation of AMPK by Ca2+/calmodulin-­dependent
­nuclear-encoded mitochondrial proteome (FIG. 2). protein kinase kinase‑β (CaMKKβ) 25 — and the
Calcium buffering The expression of nuclear-encoded mitochondrial myocyte-­specific enhancer factor 2A, promoting mito‑
A mechanism that regulates proteins is predominantly mediated by two nuclear tran‑ chondrial bio­genesis. Cold stress also activates PGC1α
calcium ion (Ca2+) scription factors: nuclear respiratory factor 1 (NRF1) through cAMP-­dependent protein kinase (also known
concentrations within and GA‑binding protein‑α (GABPα; also known as as protein kinase A (PKA)) and the cAMP response
the cytoplasm.
NRF2α). Both transcription factors, but mainly NRF1, ­element-binding protein (CREB) (FIG. 2).
Oxidative phosphorylation control the expression of genes encoding cytochrome c In addition to its role in homeostasis, anterograde
(OXPHOS). A biochemical and the vast majority of nuclear-encoded subunits that communication can also signal the existence of nuclear
pathway within mitochondria are involved in oxidative phosphorylation (OXPHOS), stress to mitochondria, thereby bringing about a reduc‑
that generates ATP through
as well as proteins implicated in mtDNA replication, tion in mitochondrial metabolism to attenuate the
the oxidation of nutrients.
transcription and translation. They also control the consequences of nuclear DNA damage. For instance,
Nuclear receptors expression of mitochondrial ribosomal proteins and artificially inducing telomere dysfunction in mice is
Ligand-dependent tRNA synthetases, rate-limiting haem biosynthetic associated with impaired mitochondrial function and
transcription factors that are enzymes, ion channel proteins and various compo‑ diminished biogenesis through the repression of PGC1α-
characterized by a central
DNA-binding domain and
nents of the protein import machinery 5–8. The expres‑ and PGC1β‑driven programmes by p53  (REF.  26) .
a carboxy-terminal sion of mitochondrial-encoded proteins also depends Activation of poly(ADP-ribose) polymerase 1 (PARP1)
ligand-binding domain. on nuclear regulation, as all of the factors that activate and PARP2 in response to DNA strand breaks likewise

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Exercise, Cold Exercise

Energetic stress responses. The retrograde stress
Ca2+
CR response was initially described in yeast, in which it
regulates carbon and nitrogen metabolism31. Yeast
PARPi AMPK CAMKKβ cAMP Ca2+ contains the retrograde response genes RTG1, RTG2
and RTG3, which induce different metabolic enzymes
DNA
and thereby activate alternative metabolic pathways to
PARP1 NAD NR PKA counteract mitochondrial dysfunction32; these include
damage
peroxisomal anaplerotic reactions such as the glyoxy-
SIRT1 NCOR1 late cycle33. Independent of the induction of the RTG
CREB CAMKIV Calcineurin genes, other pathways, including the carbon catabolite-­
RIP140
derepressing protein kinase (also known as Snf1; the
p53 PGC1α yeast orthologue of AMPK) pathway, the TORC1
Telomere pathway and the Sir2 pathway, can also be activated in
dysfunction
response to energetic stress in yeast to prevent abnormal
histone deacetylation and reprogramme metabolism,
PGC1α PGC1α PGC1α Calcineurin to regulate protein synthesis, and to improve protein
PPARs NRFs ERRs MEF2 homeostasis, respectively 34–36.
Although the absence of clear RTG orthologues
in other species hindered the simple generalization
• Mitochondrial biogenesis
• OXPHOS of the retrograde signal, some pathways are acti‑
• TCA vated in a similar fashion after mitochondrial stress,
• Fatty acid oxidation
Nucleus testify­ing to functional conservation. In C. elegans,
mutations in the ETC subunit-encoding genes iron–
Figure 2 | Anterograde regulation. Different extracellular stimuli
Nature Reviews can regulate
| Molecular Cell Biology
mitochondrial function through nuclear regulation. Exercise and caloric restriction (CR) sulfur protein 1 (isp‑1) and clk‑1 activate AMPK sub‑
activate AMP-activated protein kinase (AMPK), which, in turn, activates sirtuin 1 (SIRT1) unit α2 (REF. 37), which transduces changes in cellular
through the regulation of NAD+ levels. NAD+ can also be modulated by NAD+ precursors energy levels resulting from mitochondrial stress38.
such as nicotinamide riboside (NR) or by inhibitors of poly(ADP-ribose) polymerase 1 Defects in the TCA cycle also trigger the expression
(PARP1), termed PARPi, which are activated upon DNA damage. SIRT1 activates PPAR-γ of glyoxy­late cycle genes such as gei‑7 — an isocitrate
co-activator 1α (PGC1α), which, through interactions with peroxisome proliferator- lyase/malate synthase — through mechanisms that are
activated receptors (PPARs), nuclear respiratory factors (NRFs) or oestrogen-related ill-defined but that are most likely to involve AMP-
receptors (ERRs), induces the expression of mitochondrial genes. PGC1α signalling is activated kinase 2 (aak‑2), mediating the transition
antagonized by the co-repressors nuclear receptor co-repressor 1 (NCOR1) and to mitochondria-independent energy production39,40.
receptor-interacting protein 140 (RIP140), or by p53 in response to telomere dysfunction.
Administration of α‑ketoglutarate — a key TCA cycle
Cold and exercise can also activate the expression of mitochondrial genes by activating
PGC1α through the cAMP–protein kinase A (PKA)– cAMP response element-binding intermediate — also mediates cellular adaptations,
protein (CREB) pathway, or through Ca2+ signalling by Ca2+/calmodulin-dependent leading to lifespan ­extension through TOR, aak‑2 and
protein kinase kinase‑β (CAMKKβ) or Ca2+/calmodulin-dependent protein kinase type IV daf‑16 (REF. 41).
(CAMKIV). Additionally, Ca2+ induces mitochondrial gene expression by activating In mammals, the retrograde response has also
myocyte-specific enhancer factor (MEF2) through the phosphatase calcineurin. been linked to mTOR and AMPK; however, as mam‑
OXPHOS, oxidative phosphorylation; TCA, tricarboxylic acid. malian cells lack the glyoxylate cycle, they use differ‑
ent anaplerotic reactions to adapt their metabolism to
manage energy deficits. A decrease in mito­chondrial
represses mitochondrial function by depleting cellular ATP synthesis stimulates AMPK, promoting the acti‑
NAD+ levels27 and consequently inhibiting the activity of vation of PGC1α, which stimulates mitochondrial
SIRT1 (REF. 28). Treatment with NAD+ precursors, such energy metabolism and biogenesis23 (FIG. 3). AMPK
as nicotinamide riboside, or with PARP inhibitors can also triggers the mitochondrial quality control system,
Metabolic reprogramming
The set of changes in cellular restore NAD+ levels and activate an anterograde signal which regulates mitochondrial dynamics and induces
metabolism that allow through SIRT1–PGC1α29 (FIG. 2). mitophagy42. Similarly, reduced mTOR activity, such
quiescent cells to proliferate; as under energetic stress (for example, during exer‑
it is considered to be a Retrograde response cise and nutrient limitation), facilitates mitochondrial
hallmark of cancer.
Mitochondria generate a wide range of retrograde retro­grade signalling, whereas its activation inhibits
Reactive oxygen species signals, through which they regulate various cellular the retrograde response43.
(ROS). Reactive molecules and organismal activities and protect against mito‑
generated in cells by the chondrial dysfunction by activating the expression Ca2+-dependent stress responses. Mitochondria are
reduction of oxygen with a
of nuclear genes involved in metabolic reprogramming essential to regulate the levels of intracellular Ca 2+
single electron (superoxide),
two electrons (hydrogen or stress defence30. The retrograde response exists in (REF. 44). Different mitochondrial stressors, such as the
peroxide) or three electrons all organisms, but the regulation and nature of the loss of (or mutations in) mtDNA, disruption of ETC
(hydroxyl radical). pathways involved can vary. Despite the pleiotropic complexes and OXPHOS, or treatment with ionophores,
nature of the retrograde signals, these pathways can trigger the loss of mitochondrial membrane potential and
Anaplerotic reactions
Reactions that replenish
be classified into energetic stress responses, Ca2 +- the subsequent release of Ca2+ into the cytoplasm45–48.
intermediates of dependent responses and reactive oxygen species (ROS) Elevated levels of free cytosolic Ca 2+ activate the
metabolic pathways. stress responses, depending on the trigger. phosphatase calcineurin, which activates the nuclear

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PGC1α AMPK Low ATP

JNK ROS
Mitochondrion
• Mitochondrial biogenesis ETC
NFE2L2
• Mitochondrial quality control mtDNA
KEAP1
OXPHOS
mtDNA defects
dysfunction
Antioxidant response NFE2L2 Ca2+
degradation

NF-κB Loss of ∆ψm

• Calcium metabolism genes
• Glycolytic genes Calcineurin Ca2+ CAMKKβ

NFATC

Nucleus
PKC JNK p38 CAMKIV

EGR1 ATF2 CEBPδ CHOP CREB

Glyoxylate cycle
An anabolic pathway, and a
variation of the tricarboxylic
acid cycle, that converts two
acetyl-CoA molecules to
dicarboxylic acid (succinate);
this cycle is not present
in mammals.
Mitophagy
A selective form of autophagy
that is responsible for
the elimination of
defective mitochondria.
Ionophores
Molecules that bind to ions
and allow their passage
across a membrane or
a lipophilic phase.
Mitochondrial membrane
potential
An electrochemical potential
formed by chemiosmosis that
results from the proton
gradient generated across the
inner mitochondrial membrane
by the mitochondrial
respiratory chain.
Figure 3 | The retrograde response. Defects in oxidative phosphorylation (OXPHOS) and mitochondrial DNA (mtDNA),
Nature Reviews | Molecular Cell Biology
caused by damage in the electron transport chain (ETC) and by mutations, respectively, activate a retrograde response to
the nucleus that can be triggered by a decrease in the level of ATP, increased signalling by reactive oxygen species (ROS)
or the release of Ca2+ from mitochondria. Low ATP levels activate AMP-activated protein kinase (AMPK), which stimulates
mitochondrial biogenesis and quality control. An increase in ROS also activates anterograde regulation through AMPK
or the c‑Jun N‑terminal kinase (JNK) pathway, by activating PPARγ co-activator 1α (PGC1α). Increased levels of ROS inhibit
the kelch-like ECH-associated protein 1 (KEAP1)‑mediated proteasomal degradation of nuclear factor erythroid 2‑related
factor 2 (NFE2L2) and facilitates the translocation of NFE2L2 to the nucleus and the subsequent activation of an
antioxidant response. Loss of the mitochondrial membrane potential (Δψm) results in the release of Ca2+ from
mitochondria, inducing the expression of genes for calcium metabolism and glycolysis through two mechanisms. First,
calcineurin translocates to the nucleus with nuclear factor-κB (NF‑κB), which can also be activated by ROS, and with
nuclear factor of activated T cells (NFATC). Alternatively, Ca2+ can activate several kinases, such as protein kinase C (PKC),
JNK–p38 and Ca2+/‌calmodulin-dependent protein kinase type IV (CAMKIV), which, in turn, activate different transcription
factors such as early growth response protein 1 (EGR1), ATF2, cAMP response element-binding protein (CREB),
CCAAT/enhancer-binding protein‑δ (CEBPδ) and CEBP homologous protein (CHOP). The release of Ca2+ can also activate
anterograde regulation through Ca2+/calmodulin-­dependent protein kinase kinase‑β (CAMKKβ).
factor‑κB (NF‑κB) p105 subunit and nuclear factor ROS-dependent responses. Mitochondrial ROS are
of activated T cells (NFATC)49–51 (FIG. 3). Both of these gener­ated through aerobic metabolism, often result‑
transcription factors then translocate to the nucleus, ing from defective ETC, and they function directly by
where they promote the synthesis of proteins involved regulating redox biology and as signalling molecules
in Ca 2+ transport and storage 50,52, and of glycolytic in numerous cellular processes, under normal as well
and gluconeogenic enzymes 52. Elevated Ca 2+ levels as stress conditions55,56. In C. elegans, mutation in clk‑1
also directly activate different Ca2+-regulated kinases, — a mitochondrial hydrolase that is required for syn‑
such as CAMKIV, Ca2+-dependent protein kinase C, thesizing the ETC component ubiquinone57 — inhibits
c‑Jun N-terminal kinase (JNK) and p38 MAPK; respiration and increases ROS levels, which stabilize
these, in turn, stimulate different transcription fac‑ and activate hypoxia-inducible factor 1 (HIF‑1), a tran‑
tors such as CREB, early growth response protein 1 scription factor that regulates adaptation to low oxy‑
(EGR1), cAMP-dependent transcription factor ATF2, gen levels58. ROS also signal through the orthologue of
CCAAT/enhancer-binding protein‑δ (CEBPδ) and stress-­inducible p38 MAPK, PMK‑1, which phosphory­
CEBP homologous protein (CHOP)45,49 (FIG. 3). Which lates and activates the transcription factor skinhead 1
of these transcription factors is activated depends on (SKN‑1), thereby promoting its nuclear localization59–61.
the cell type and the activating stimulus; furthermore, These mechanisms induce the expression of adaptive
activation not only facilitates mitochondrial adaptation ROS defence genes, such as superoxide dismutase and
but also leads to pleiotropic responses affecting calcium catalase60. CLK‑1 itself can also function as a messen‑
metabolism, insulin signalling, glucose metabolism ger of mitochondrial stress, as certain CLK‑1 isoforms
and cell proliferation24,25,48–54. translocate to the nucleus following ROS formation

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Mitochondria fuel switching
Adaptations in mitochondrial
function and electron transport
chain organization as a result of
changes in the availability
of the fuel source.
Mitochondrial uncoupling
The dissociation of
mitochondrial respiration from
ATP generation, characterized
by increased permeability of
the inner mitochondrial
membrane to protons and
subsequent dissipation of
mitochondrial
membrane potential.
Mitonuclear imbalance
Mitonuclear imbalance occurs
when oxidative
phosphorylation subunits
encoded by both
mitochondrial and nuclear
DNA fail to assemble together
in precise stoichiometric ratios
to ensure proper mitochondrial
function, owing to an increase
or decrease of subunits from
one of the origins.
and induce a protective programme, while inhibiting
the mitochondrial unfolded protein response (UPRmt)
by binding chromatin, a property that is conserved in
mammalian cells62.
In Drosophila melanogaster, disruption of complex I of
the ETC triggers a signalling cascade that involves ROS-
mediated dimerization of the D. melanogaster homologue
of the mammalian kinase ASK1, which promotes Jnk
signalling to the forkhead box O (Foxo) transcription
factor and activates the cyclin-dependent kinase inhib‑
itor Dacapo63. In parallel, increased AMP levels arising
from a decrease in mitochondrial ATP synthesis activate
AMPK and p53, leading to the down­regulation of cyclin
E1 and subsequent G1 arrest as part of the G1–S cell cycle
checkpoint63. Through the Jnk pathway, ROS also activate
the UPRmt in flies with muscle-­specific depletion of the
complex I subunit NADH–ubiquinone oxidoreductase
75 kDa (Nd75)64 (see below).
In mammals, increases in ROS to levels that are not
deleterious to cell function induce a retrograde signal to
activate detoxification enzymes and antioxidant proteins
in mitochondria and the cytosol65–68. This activation is
mediated by the binding of transcription factors such as
nuclear factor erythroid 2‑related factor 2 (NFE2L2; also
known as NRF2, but not to be confused with GABPα
mentioned above) to antioxidant response elements69
(FIG. 3). Increased levels of mitochondrial ROS were also
reported to activate NF‑κB, thereby promoting cellular
proliferation and survival in cancer cells51. ROS can also
induce mitochondrial biogenesis and the expression of
genes involved in OXPHOS by promoting JNK–PGC1α
signalling 70, and can promote mitochondria fuel switching
by mediating complex II phosphorylation71 and meta‑
bolic reprogramming through mitochondrial uncoupling
and AMPK activation72.
Mitonuclear feedback and proteostasis
A well-conserved protein quality control system, com‑
prising mainly chaperones and proteases encoded in
the nucleus, exists in mitochondria to maintain mito­
chondrial proteostasis73. These proteins participate in
the folding, assembly and turnover of mitochondrial
proteins under both normal and stress conditions.
Stresses that exceed the capacity of this protein quality
control system are sensed by mitochondria and com‑
municated to the nucleus to promote the expression of
these quality control components, as well as other com‑
pensatory genes that have been implicated in restoring
mito­chondrial homeostasis through bidirectional com‑
munication between the two organelles. Depending on
the nature of the stressor, three different mitonuclear
proteostasis responses have been described: the UPRmt,
which activates the expression of proteases, chaper‑
ones and other stress response genes; proteolytic stress
responses, which specifically induce the expression
of some mitochondrial proteases; and the heat shock
response, which activates mitochondrial chaperones.
The UPRmt. The UPRmt is a protective transcriptional
response that promotes the expression of mitochondrial
proteostasis genes to stabilize mitochondrial function,
and of metabolic genes to adapt to the provoking stress.
The UPRmt is triggered by mitochondrial proteotoxic
stresses, such as the accumulation of unfolded proteins,
impairment of the protein quality control system, mito-
nuclear imbalance or inhibition of the ETC74. The UPRmt
has been characterized most fully in C. elegans, in which
it can be induced using RNA-mediated interference
(RNAi) directed against some nuclear-encoded mito‑
chondrial proteins, such as cytochrome c oxidase (cco‑1),
a mitochondrial protein quality-control protease, spastic
paraplegia 7 (spg‑7); or several mitochondrial ribosomal
proteins, typified by mitochondrial ribo­somal protein S5
(mrps‑5)75,76 (see below for more details about mrps‑5).
Furthermore, doxycycline and chloram­phenicol —
which are antibiotics that target bacterial as well as
mitochondrial translation owing to the evolutionary
conservation of the machinery — activate the UPRmt by
depleting mitochondrial-encoded ETC subunits, simi‑
lar to mtDNA depletion by ethidium bromide75. PARP
inhibitors or NAD+ precursors that activate SIRT1 and
mitochondrial function, as well as the longevity com‑
pounds resveratrol and rapamycin, also activate the
UPRmt owing to a mitonuclear imbalance, which in
this case is generated by enhancing the production of
mtDNA-encoded ETC proteins77–79.
The UPRmt in C. elegans involves the digestion by the
matrix protease CLPP of unfolded or unassembled mito‑
chondrial proteins into peptides, which are transported
into the cytoplasm by the HAF‑1 transporter 80 (FIG. 4a).
Through a poorly understood mechanism, the cyto‑
plasmic accumulation of these peptides induces a tran‑
scriptional response coordinated by ATFS‑1 (activating
transcription factor associated with stress 1). ATFS‑1
possesses a mitochondrial targeting sequence as well as
a nuclear localization signal. Under normal conditions, it
constantly shuttles to mitochondria, where it is degraded
by the LON protease (LONP); however, in response to
mitochondrial stress, ATFS‑1 import to mito­chondria
is attenuated, causing it to localize in the nucleus
instead81. Here, ATFS‑1 and two other factors, DVE‑1
and ubiquitin-­like 5 (UBL‑5), induce the expression of
several genes that are involved in mitochondrial quality
control and cellular metabolism to restore proteostasis.
These include the mitochondrial chaperones heat shock
protein 6 (hsp‑6), hsp‑60 and DNaJ domain 10 (dnj‑10);
the intermembrane ATPases associated with diverse
cellu­lar activities (iAAA) protease ymel‑1; the mito‑
chondrial fission factor drp‑1; glycolytic genes such as
gpd‑2; detoxification genes such as the transcription fac‑
tor skn‑1; and the core components of the translocase of
the inner membrane 23 (TIM23) complex, tim‑23 and
tim‑17 (REFS 80–84) (FIG. 4a). During stress, ATFS‑1 also
limits the expression of other mitochondrial genes in the
nucleus, such as those encoding TCA cycle enzymes and
ETC subunits, by repressing their promoters84. To ensure
that the expression of mitochondrial and nuclear ETC
subunits is coordinated under stress conditions, cer‑
tain splice variants of ATFS‑1 are specifically imported
into mitochondria during stress (through an unknown
mechanism), where they repress the expression of
mtDNA-encoded ETC subunits84 (FIG. 4a).

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The UPRmt has also been described in D. melano‑ an internal deletion (ΔOTC), which causes mitochon‑
gaster, although it has been less extensively studied in drial protein overload, induces the expression of the
this model. Overexpression of a mutated form of the D. melanogaster orthologues of the chaperones HSP60
mitochondrial ornithine transcarbamylase containing and mtHSP70 (REF. 85). Mild mitochondrial distress in

a Caenorhabditis elegans
Mitochondrion Nucleus
? • OXPHOS
OXPHOS ?
ATFS-1 ? ATFS-1 • TCA genes
ATFS-1
mtDNA

? • Glycolysis genes
LONP ? ATFS-1
ATFS-1 • OXPHOS assembly

CLPP ? DVE-1
HAF-1 DVE-1 Mitochondrial
UBL-5 ATFS-1 UBL-5 chaperones
and proteases

b Mammals
UPRmt • CHOP
JNK2 JUN • CEBPβ

CEBPβ
CHOP
Mitochondrial
chaperones
and proteases
• HSP60 MURE CHOP MURE
• HSP10
• CLPP
? • Mitophagy
SIRT3 FOXO3A FOXO3A • ROS detoxification

Proteolytic stress responses ? • LONP

• AFG3l2
• SPG7
StAR StAR • YME1l1

Heat stress response HSF1 BRG1 Mitochondrial

SSBP1 SSBP1 SSBP1 HSF1 and cytosolic
chaperones

Figure 4 | Mitonuclear feedback. a | In worms, the CLPP-mediated cleavage of unfolded proteins in mitochondria initiates
Nature Reviews | Molecular Cell Biology
mitochondrial unfolded protein response (UPRmt) signalling. The efflux of short peptides through the mitochondrial
transporter HAF‑1 somehow inhibits mitochondrial protein import and, consequently, the transcription factor ATFS‑1
(activating transcription factor associated with stress 1), which is normally imported into mitochondria and degraded by the
LON protease (LONP), translocates into the nucleus. Here, together with ubiquitin-like 5 (UBL‑5) and DVE‑1, it induces
the expression of UPRmt target genes — namely, those encoding chaperones and proteases — to restore proteostasis.
ATFS‑1 can also positively regulate glycolysis and assembly of the oxidative phosphorylation (OXPHOS) system and
negatively regulate the expression of tricarboxylic acid (TCA) cycle and OXPHOS genes. In response to mitochondrial stress,
splice variants of ATFS‑1 accumulate in mitochondria and repress the expression of mitochondrial OXPHOS genes.
b | In mammals, the activation of JUN by c-Jun N-terminal kinase 2 (JNK2) leads to the induction of CEBP homologous protein
(CHOP) and CCAAT/enhancer-binding protein-β (CEBPβ), which heterodimerize and activate UPRmt genes. Mitochondrial
proteotoxic stress also activates sirtuin 3 (SIRT3), which deacetylates forkhead box transcription factor FOXO3A, causing its
translocation to the nucleus to induce the expression of reactive oxygen species (ROS) detoxification and mitophagy genes.
The import of steroidogenic acute regulatory protein (StAR) from the mitochondrial outer membrane into the matrix
induces, through an unknown mechanism, the expression of several proteases, which subsequently clear StAR from
mitochondria. Heat causes single-stranded DNA-binding protein 1 (SSBP1) to translocate into the nucleus, in association
with heat shock factor 1 (HSF1), to bind the chromatin modifier BRG1 and induce the expression of mitochondrial
and cytosolic chaperones. AFG3L2, AFG3‑like protein 2; HSP, heat shock protein; mtDNA, mitochondrial DNA;
MURE, mitochondrial unfolded response element; SPG7, spastic paraplegia 7; YME1L1, YME1-like protein 1.

218 | APRIL 2016 | VOLUME 17 www.nature.com/nrm

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Mitohormesis
A process in which low levels of
mitochondrial stress cause a
protective cellular response
that reduces susceptibility to
disease and potentially
promotes longevity.
Mitofusins
High-molecular-mass GTPases
that are essential for
mitochondrial fusion.
Mouse BXD genetic
reference population
A family of recombinant inbred
mouse strains, which is
currently the largest and
best-characterized mouse
genetic reference population,
composed of ~160 lines.
muscle cells arising from the muscle-specific knockdown
of Nd75 from complex I in D. melanogaster also induces
the UPRmt, together with a mitohormesis response64
(see below).
Several lines of evidence suggest a certain degree of
conservation of the existence of the UPRmt in vertebrates,
including in humans. The first line of evidence comes
from studies using cell lines. In monkey kidney COS7
cells, the mitochondrial protein overload generated by
ΔOTC causes the activation of CHOP and CEBPβ86,87.
Heterodimers of these transcription factors bind to con‑
served mitochondrial unfolded response elements and
induce the expression of UPRmt genes, including those
encoding the chaperones HSP60, HSP10 and mtDNAJ;
the proteases YME1L1, CLPP and PMPCB; the mito‑
chondrial import complex subunit TIM17A; and some
mitochondrial enzymes, such as thioredoxin 2 (REF. 88).
Knockdown of mitochondrial Leu-rich PPR motif-­
containing protein, which impairs complex IV, acti‑
vates the UPRmt by inducing mitonuclear imbalance in
a neuroblastoma cell line and in worms89. Independent
of CHOP, the mammalian mitochondrial deacetylase
SIRT3 is also activated following mitochondrial proteo‑
toxic stress; SIRT3 coordinates an antioxidant response
and mitophagy 90 (FIG. 4b). Overexpression of CLPX, the
regulatory subunit of complex CLPXP, is reportedly
sufficient to induce the UPRmt in C2C12 myoblasts91.
Inhibition of HSP90 chaperones in the mitochondria of
a human glioblastoma cell line triggers the UPRmt and
autophagy, potentially by inducing CHOP and CEBPβ
and repressing NF-κB92. Besides the induction of UPRmt,
over­expression of ΔOTC also leads to the activation of
mitophagy, suggesting communication between the
­different ­mitochondrial quality control systems93.
Further evidence comes from several mouse models
with mitochondrial dysfunction, in which the UPRmt is
induced, indicating that it seems to exert homeostatic
functions in mammals. For instance, deficiency of aspar‑
tyl tRNA synthetase in mouse skeletal and heart muscle
causes the loss of mitochondrial proteostasis, which, in
the heart, activates the UPRmt and promotes mitochon‑
drial biogenesis94. Cardiomyocyte-specific deletion of
the mitofusins in mice prevents mitochondrial fusion,
thereby impairing mitophagy and causing the accumu‑
lation of dysfunctional mitochondria and activation of
the UPRmt (REF. 95). A mutation in the gene encoding the
mitochondrial helicase Twinkle, which causes multiple
mtDNA deletions and late-onset mitochondrial disease
in mice, induces a progressive OXPHOS deficiency
accompanied by an increase in HSP60, mtHSP70 and
CLPP protein levels in muscle96. The histone deacetylase
SIRT7, which has emerged as a master regulator of
mitochondrial homeostasis97, can also promote, in con‑
junction with NRF1, the regenerative capacity of aged
haematopoietic stem cells through a mechanism that
potentially involves the UPRmt (REF. 98). Finally, another
indication in support of conservation of the UPRmt in
mammals comes from studies in the mouse BXD genetic
reference population, in which the expression of the proto­
typical UPRmt gene network, comprising mammalian
orthologues of six worm UPRmt regulators, negatively
correlates with the expression of Cox5b/COX5B and
Spg7/SPG7 — the mouse/human orthologues of worm
cco‑1 and spg‑7 — suggesting that the low abundance of
these genes triggers the UPRmt in mammals as it does
in C. elegans 99.
Proteolytic stress responses. In certain contexts, mito‑
chondrial proteotoxicity can lead to a specific proteolytic
response and, in the case of the accumulation of unfolded
proteins within the intermembrane space, genes other
than those involved in the canonical UPRmt are activated
to resolve the stress. Overexpression of a mutant form
of endonuclease G reportedly causes stress in the inter­
membrane space, leading to the production of ROS and
phosphorylation of AKT (also known as protein kinase
B)100; consequently, the intermembrane space Ser protease
HTRA2 (also known as OMI) and NRF1 are induced,
­ultimately leading to increased proteasome activity 100.
The steroidogenic acute regulatory protein (StAR)
overload response (SOR) is another form of proteo‑
lytic stress and occurs in the context of the regulation
of steroido­genesis in the adrenal cortex and gonads101.
StAR contributes to steroid synthesis by transferring
cholesterol through the outer mitochondrial mem‑
brane to the inner mitochondrial membrane, where it is
converted into steroids. StAR accumulation and activ‑
ity are prevented by its import into the mitochondrial
matrix 102, which is followed by sequential degradation by
­mitochondrial proteases101 (FIG. 4b).
Heat shock response. Heat stress constitutes a threat to
proteins in all cellular compartments103, but it can also
elicit a specific mitochondrial signal to the nucleus,
causing the expression of the mitochondrial chaperones
HSP10 and HSP60, in addition to regular heat shock pro‑
teins104. In cultured cells, heat causes the translocation of
the mitochondrial single-stranded DNA-binding protein 1
(SSBP1) in a complex with heat shock factor 1 (HSF1)
into the nucleus, where, by recruiting the chromatin-­
remodelling factor BRG1, it induces the expression of
mitochondrial and cytoplasmic–nuclear chaperones that
are essential for maintaining the mitochondrial membrane
potential and for survival following heat shock (FIG. 4b).
Integrated stress response
Mitochondrial stress can also activate the ISR, which is
a general cellular response that modulates global pro‑
tein synthesis (FIG. 5). This response is induced by dif‑
ferent stress stimuli, including the ER unfolded protein
response (UPRer), the UPRmt, oxidative stress, nutrient
deprivation, the presence of viral double-stranded RNA
or haem deficiencies105,106. The key component of this
stress response is the α-subunit of eukaryotic trans­lation
initiation factor 2 (eIF2α). Phosphorylation of eIF2α
by several different kinases — including general con‑
trol non-derepressible 2 (GCN2), PKR-like ER kinase,
protein kinase double stranded RNA-dependent or
haem-regulated inhibitor — globally inhibits protein
synthesis but concurrently facilitates the specific expres‑
sion of stress-response genes107 such as ATF4 (REF. 108).
ATF4, in turn, induces the expression of a wide range

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Mitochondrion Nucleus

Amino acid ER Haem deficiency,

starvation stress dsRNA heavy metals
ETC
• CHOP
ROS GCN2 PERK PKR HRI • GADD34
• ATF3
ATF4 • TRIB3
P
eIF2α eIF2α • BIP

Cell survival Apoptosis

Protein Translation of
translation ATF4

Cytosolic
mPOS, homeostasis
UPRam genes
MTS
UPRam

Proteasome

Figure 5 | The integrated stress response. The central regulation node of the integrated stress response (ISR) is the
eukaryotic translation initiation factor eIF2α, which, when phosphorylated, inhibitsNature Reviews
cytosolic | Molecular
translation. Cell Biology
Four different
kinases are known to phosphorylate eIF2α in response to various stresses: general control non-derepressible 2 (GCN2) is
activated by amino acid starvation; PKR-like ER‑kinase (PERK) is activated following endoplasmic reticulum (ER) stress;
protein kinase double stranded RNA-dependent (PKR) is activated by double-stranded RNA (dsRNA) following viral
infection; and haem-regulated inhibitor (HRI) is activated by heavy metals and haem deficiency. Defects in the electron
transport chain (ETC), reactive oxygen species (ROS) and mitochondrial proteotoxic stresses can activate GCN2, PERK or
HRI, depending on the context. Phosphorylation of eIF2α promotes the selective translation of the transcription factor ATF4,
which, in turn, promotes the expression of CEBP homologous protein (CHOP), growth arrest and DNA damage-inducible
protein 34 (GADD34), ATF3, tribbles homologue 3 (TRIB3) and immunoglobulin heavy chain-binding protein (BIP), as well as
other transcription factors, to restore cellular homeostasis. However, apoptosis can also ensue in the case of irreversible
cellular damage. In yeast, under conditions of mitochondrial stress, when protein import to mitochondria is slowed, newly
translated mitochondrial polypeptides accumulate in the cytosol, inducing mitochondrial precursor over-accumulation
stress (mPOS). This accumulation blocks translation and triggers an unfolded protein response activated by mis-targeting of
proteins (UPRam), which activates the proteasomal degradation pathway and induces the expression of gene sets that are
involved in restoring cellular homeostasis. MTS, mitochondrial targeting sequence.

Mitokines
Signals released from a cell or
tissue in response to
mitochondrial stress to
modulate cellular or
organismal homeostasis
and longevity.
Ketogenesis
A metabolic pathway in the
liver that generates ketones
using acetyl-CoA, which
provides an important fuel
source for the brain and other
tissues during long-term
fasting.
of stress proteins, such as CHOP, growth arrest and
DNA damage-­inducible protein 34 (GADD34), ATF3,
immuno­globulin heavy chain-binding protein (BIP; also
known as GRP78) or tribbles homologue 3 (TRIB3), to
restore proper c­ ellular function105,109,110.
The ISR and the phosphorylation of eIF2α as its pivot
are highly conserved. In C. elegans, mutations in several
ETC genes, such as clk‑1 or isp‑1, induce ROS-mediated
phosphorylation of eIF2α by the kinase GCN‑2, thereby
reducing cytosolic translation and the generation of new
mitochondrial proteins111. Activation of the ISR in this
context seems to be crucial, as loss of function of GCN2
affects mitochondrial activity and sensitizes cells to
mitochondrial stress, with detrimental consequences111.
In mammals, different mitochondrial stressors can
activate the ISR. Arsenic-mediated mitochondrial
stress, through eIF2α phosphorylation, impairs the
function of the mitochondrial protein import complex
and activates a gene expression programme for main‑
taining mitochondrial proteostasis in mammals, as
well as in C. elegans 112. Altered or impaired expression
of mtDNA in different mammalian cell types activates
the GCN2‑mediated ISR, triggering the expression of
several different stress proteins, such as CHOP, TRIB3,
ATF3 and hairy/enhancer-of‑split related with YRPW
motif protein113. Tetracyclines such as doxycycline,
which induce mitonuclear imbalance and mitochondrial
dysfunction75,114, can also trigger the ATF4‑mediated
expression of stress genes 115. The accumulation of
unfolded proteins in the inner mitochondrial membrane
that is generated by the loss of HTRA2 in the brain also
activates a CHOP-dependent ISR, protecting against
neuronal cell death116. In cultured cells, the outcome of
the link between mitochondrial stress and the ISR can
be protective or deleterious, probably depending on the
intensity and duration of the mitochondrial insults. In
certain cells, inhibition of some mitochondrial ETC
subunits activates an ATF4‑dependent ISR, promot‑
ing cell viability and metabolic reprogramming 117,118;
however, the mitochondrial-dependent activation of
the ISR is deleterious in other cell types, inhibiting cell
­proliferation or altering lipid metabolism119,120.
Finally, beyond the prototypical ISR, other mech­
anisms that control and decrease global protein synthe‑
sis can be activated under conditions of mitochondrial
stress, especially in yeast. In this organism, mito­
chondrial stress generated by loss of mitochondrial
membrane potential or by defects in proteins of the mito‑
chondrial import machinery decreases mito­chondrial
protein import, resulting in the aberrant cytosolic

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accumulation of newly synthesized unfolded mitochon‑ Mitochondrial stress modulates lifespan

drial peptides121,122. This so‑called mitochondrial precur‑
sor over-accumulation stress (mPOS) can not only lead
to mitochondria-mediated cell death, but can also miti­
gate the stress by upregu­lating a large network of cyto‑
solic genes to promote alternative pathways of protein
synthesis and folding, as well as cell survival121 (FIG. 5).
The mPOS response can also induce a UPR activated
by mis-targeting of proteins (UPRam), which protects the
cell against defects in mitochondrial import by ­inhibiting
protein synthesis and activating the proteasome122.
Stress and extracellular communication
Mitochondrial stress signals not only activate different
cellular responses but also trigger non-cell-autonomous
responses, implying that mitokines can be secreted from
cells with stressed mitochondria. In C. elegans, mito‑
chondrial stress triggered by knockdown of ETC sub‑
units in a tissue-specific manner can be transduced to
distant tissues: when mitonuclear signalling was selec‑
tively induced in neurons by neuron-specific knock‑
down of cco‑1, the UPRmt was not only activated in the
nervous system but also in the intestine, with a beneficial
effect on the whole organism, including an increase in
lifespan76. The origin of these inter-tissue cues is crucial
for longevity, as the beneficial effects of UPRmt activa‑
tion are lost if the cues do not originate in the neurons76.
Similarly, in D. melanogaster, knockdown of ETC-
encoding genes in neurons enhances longevity, implying
the existence of systemic communication of an unknown
nature that affects the whole organism123. Likewise, mild
ETC deficiency in fly muscles is communicated to other
tissues through insulin signalling, which confirms the
non-cell-autonomous nature of the signal64.
In mammals, the mitochondria-mediated
non-cell-autonomous response is thought to be regu­
lated either by mitochondria-derived peptides or by
cellular hormones. Although different mitochondria-­
derived secreted peptides have been described, further
mechanistic studies are needed to identify their exact
modes of action. Some mitochondria-derived peptides
are encoded by open reading frames identified in the
mtDNA genome124. For example, the peptide humanin,
which is encoded by the 16S rRNA gene, is secreted from
mitochondria and was reported to have cyto­protective
properties in a model of Alzheimer disease125,126. Another
mitochondria-derived peptide is encoded within the
sequence encoding mitochondrial 12S rRNA and pro‑
motes AMPK activation in the muscle, thereby regu‑
lating metabolic homeostasis and protecting against
insulin resistance and obesity 124. Fibroblast growth
factor 21 (FGF21) is a fasting-regulated hormone with
pleiotropic actions in different tissues127. Interestingly,
FGF21 functions as a potent mitokine and is secreted
from skeletal muscle into the circulation in humans and
mice with deficiencies in OXPHOS, in a manner medi‑
ated by the ATF4‑dependent ISR128–130. When it reaches
the liver, FGF21 enhances ketogenesis and protects
Rho0 cells
against diet-­induced obesity and insulin resistance by
Eukaryotic cells that lack enhancing β‑oxidation, lipolysis and browning of white
mitochondrial DNA. adipose tissue130.
Mitochondrial dysfunction constitutes a hallmark of
and a driving force behind ageing 131, and mitochondrial
insults underpin the development of several diseases132.
Conversely, subjecting mitochondria to specific stresses
extends lifespan across species in a manner dependent
on mitonuclear communication. The most common
lifespan extender is attenuation of OXPHOS, which
decreases respiration and concomitantly increases
lifespan in different organisms (TABLE 1), although it is
often associated with slower development, reduced body
size and infertility 133,134. Several different, non-mutually
exclusive mechanisms have been proposed to explain
mitochondrial stress-dependent lifespan extension,
including the retrograde stress response activated by
energy deficiency, ROS135 and Ca2+ signalling, as well as
by the UPRmt (REF. 76).
Energy regulation. In yeast, activation of the retro‑
grade response increases chronological lifespan, and
RTG2 and RTG3 are required to increase the chrono‑
logical and replicative lifespan of respiration-deficient
strains136. Other retrograde pathways in yeast, such as
those involving TORC1 and transcription factor Sfp1
pathways, and the activation of Sir2, also increase rep‑
licative lifespan, independently of RTGs35,36. Activation
of AMPK is required to extend the chronological
lifespan of rho 0 cells 34. Consistent with its negative
effect on the retrograde response, disruption of TOR
signalling without a preceding mitochondrial stress
also increases yeast replicative lifespan137. AMPK is also
required for the increased longevity that is induced in
the worm isp‑1 and/or clk‑1 mutants37,138, as well as
for the lifespan extension that is seen following loss of
function of the worm p53 homologue cep‑1 (REF. 139) or
putative homeobox protein 23 (REF. 140). Retrograde
signalling through skn‑1 also governs longevity,
coordinat­ing the bimodal regulation of mitochondrial
bio­genesis and mitophagy 141. Alterations in the TCA
cycle or supplementation with some of its intermedi‑
ates such as α‑ketoglutarate l­ ikewise increase longevity
in worms39,41 (TABLE 1).
ROS signalling. Moderate levels of ROS are also c­ rucial
transducers of mitochondrial stress that induce longev‑
ity in a range of organisms142. Following inhib­ition of
respiration, ROS activates HIF‑1 (REF. 58) and increases
lifespan in C. elegans 143. skn‑1 is also pivotal in trans­
ducing longevity cues that involve ROS signalling 60.
Low doses of toxic compounds that generate ROS,
such as paraquat, arsenite or juglone, consistently
extend C. elegans lifespan61,144,145. This pro-longevity
effect depends specifically on mitochondrial ROS, as
elevated levels of cytosolic ROS shorten lifespan146.
ROS signalling is also required to induce longevity
mediated by other types of mitochondrial communi‑
cation: ROS are necessary for UPRmt cell-autonomous
signalling in flies lacking ND75 in muscle cells and for
their increased longevity, whereas antioxidants reverse
longev­ity in some ETC worm mutants, such as clk‑1
and isp‑1 (REFS 64,135).

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Table 1 | Extended longevity triggered by mitochondrial stress

Function affected Organism Stressor Mediator or stress pathway Refs
Redox homeostasis Worm Paraquat, juglone or arsenite at low DAF‑16, SIR‑2.1, AAK‑2, ROS 61,144,
levels 145
TCA cycle Worm Malate or fumarate DAF‑16 39
α‑ketoglutarate TOR/LET‑363, AAK‑2, DAF‑16 41
Oxaloacetate AAK‑2, DAF‑16 168
ETC (complex I) Worm nuo‑2 depletion ROS 169
nuo‑6 mutation or depletion ROS 170
Fly Muscle-specific Nd75 depletion Non-cell-autonomous insulin 64
signalling
ETC (complex III) Worm cyc‑1 depletion ROS 169
isp‑1 mutation AAK‑2 133
isp‑1 depletion ROS 170
ETC (complex IV) Worm cco‑1 depletion UPRmt 76,169
cco‑1 intestinal- or neuronal-specific Non-cell-autonomous UPRmt 76
depletion
Mouse Surf1 knockout Calcium? 149
ETC (complex V) Worm atp‑3 depletion ROS 169
ETC (complexes I, II, Fly Depletion of complexes I, II, III, IV or V ? 123
III, IV or V)
Coenzyme Q Worm clk‑1 mutation ROS, HIF‑1, AAK‑2 58,134
biosynthesis
Fly Sbo (Coq2) depletion ? 171
Mouse Mclk1 heterozygosity ROS 147
Mitochondrial Worm CCCP ? 172
uncoupling
Mouse 2,4‑DNP ? 152
Mitochondrial Worm and mrps‑5 depletion or variation in Mrps5 UPRmt 75
translation mouse levels
Worm Doxycycline and chloramphenicol UPRmt 75
NAD levels
+
Worm Nicotinamide riboside SIR‑2.1, DAF‑16, UPR mt
77
Mitochondrial Fly Overexpression of Hsp22 ? 153
chaperones
Overexpression of Hsp60 in muscle ? 64
2,4‑DNP, 2,4‑dinitrophenylhydrazine; AAK‑2, AMP-activated kinase 2; atp‑3, ATP synthase subunit 3; CCCP, carbonyl cyanide m‑chloro-
phenylhydrazone; cco‑1, cytochrome c oxidase 1; cyc‑1, cytochrome c; ETC, electron transport chain; HIF‑1, hypoxia-inducible factor 1;
Hsp, heat shock protein; isp‑1, iron–sulfur protein 1; mrps‑5, mitochondrial ribosomal protein S5; nuo, NADH ubiquinone oxidoreductase;
ROS, reactive oxygen species; Surf1, surfeit gene 1; TCA, tricarboxylic acid; UPRmt, mitochondrial unfolded protein response.

Inactivation of some mouse orthologues that are
mutated in long-lived worms and flies also increases
longevity (TABLE 1) through retrograde ROS or Ca2+ sig‑
nalling. Mice heterozygous for a mutation in Mclk1 (also
known as Coq7), which encodes an enzyme involved
in coenzyme Q biosynthesis, have ETC defects and live
longer147,148. Similarly to flies, deficiency in the complex IV
assembly factor surfeit locus protein 1 (SURF1) also
increases longevity in mice149,150. The longevity induced
by mutations in Mclk1 and in Surf1 is thought to involve
retrograde signals that depend on ROS and Ca2+, respec‑
tively 147,149; however, Surf1 mutants also show an increase
in mitochondrial biogenesis, a tissue-specific induction
of the UPRmt and an antioxidant response mediated
by NFE2L2 (REF. 151). Conversely, mild mitochondrial
uncoupling, which enhances mitochondrial respiration
and decreases ROS production, also increases the lifespan
of mice by modulating their metabolic conditions152.
The UPRmt. In worms, the longevity seen in loss-of-
function cco‑1 mutants depends on UBL‑5 and the
induction of the UPRmt (REF. 76). Similarly, the extended
lifespan that is triggered by mitonuclear imbalance
induced by altered mitochondrial translation also
depends on activation of the UPRmt (REF. 75). This mech‑
anism is evolutionarily conserved, as the expression of
Mrps5 correlates inversely with longevity across the
mouse BXD genetic reference population, as it does
in C. elegans 75. C. elegans mutants of clk‑1 and isp‑1,
which also display the ATFS‑1‑dependent UPRmt, have
an extended lifespan that is dependent on activation
of the ISR111. Of note, forced nuclear localization of
clk‑1, which functions as a suppressor of UPRmt gene
expression, abrogates the longevity of clk‑1 mutants62.
UPRmt genes are activated in flies with extended lifespan
that lack Nd75 specifically in muscle cells, and forced
expression of Hsp60 in muscles is sufficient to extend

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Box 1 | Mitonuclear stress signalling and the immune response
Given their crucial functions in energy metabolism, mitochondria are the Achilles heel
of cellular activities, constituting a prime target for attacks by pathogens and toxins
that target mitochondrial activity and integrity159. Furthermore, the proto-bacterial
origins of mitochondria makes it tempting to speculate that cellular defence pathways
against pathogens have similarities with those that defend against mitochondrial
stress: when some molecules, such as mitochondrial DNA (mtDNA) and formyl peptides,
are released from damaged mitochondria, the immune system can be activated as
it is during bacterial infection. The mitochondrial unfolded protein response
(UPRmt) is involved in both mitochondrial and immune surveillance, to resolve
mitochondrial stress and infection alike. Several bacterial species encountered by
Caenorhabditis elegans induce mitochondrial dysfunction and consequently activate
the UPRmt as a protective response160. Mutants in the UPRmt genes ubl‑5
(ubiquitin-like 5), dve‑1 and atfs‑1 (activating transcription factor associated with
stress 1) display weakened immunity and reduced survival against bacteria that are
commonly pathogenic for C. elegans138,161. Following exposure to Pseudomonas
aeruginosa, ATFS‑1 induces the expression of genes encoding components of the
innate immunity response, such as lysozyme and antimicrobial peptides161. Immunity is
also intimately linked to signalling by reactive oxygen species (ROS), as ROS activate
aak‑2 (AMP-activated kinase 2) and hif‑1 (hypoxia-inducible factor 1), leading to
increased resistance of C. elegans to pathogenic strains of Escherichia coli138.
In mammals, mitochondrial stress also elicits an enhanced immune response.
Moderate mitochondrial DNA stress induced by loss of function of TFAM (transcription
factor A, mitochondrial) activates a cytosolic antiviral signal that enhances the
expression of a subset of interferon-responsive genes162. Mitochondria are also involved
in antiviral defence through the mitochondria-associated viral sensor, a mitochondrial
outer membrane protein that activates an antiviral signalling response159. Mitochondrial-
generated ROS contribute to the bactericidal activity of macrophages by activating cell
surface Toll-like receptors163. ROS also enhance immunity of mice heterozygous for a
mutation in the gene encoding mitochondrial 5‑demethoxyubiquinone hydroxylase
— a protein involved in ubiquinone biosynthesis — against Salmonella enterica and
tumour cell grafts, through the activation of HIF1α164,165. Cellular protection against
intracellular pathogens such as Mycobacterium spp. or S. enterica is furthermore
dependent on mechanisms similar to mitophagy (the removal of defective
mitochondria)166. Conversely, extreme mitochondrial stress caused by TFAM deficiency
in T cells causes lysosomal defects and an increased inflammatory response, which are
recovered by restoring NAD+ levels and mitochondrial function167.
lifespan64. Similarly, overexpression in D. melanogaster
of the chaperone Hsp22, which is induced upon oxi‑
dative stress, is sufficient to increase lifespan, which
suggests a close ­connection between longevity and
UPRmt signalling 153.
Conclusion and future perspectives
Coordination between mitochondria and the nucleus
is essential for maintaining cellular homeostasis and
ensuring an effective response to stress. Mitochondrial
function is regulated by the nucleus through several
transcription factors that activate the expression of the
mitochondrial genome; through the transcriptional
induction of several mitochondrial proteins encoded by
nuclear DNA; and by promoting mitochondrial adaptive
pathways. Similarly, mitochondria can regulate nuclear
function by controlling the levels of various metabo­
lites that affect gene expression, such as ATP, acetyl-
CoA, NAD+ and ROS154. Mitochondria maintain their
Formyl peptides homeo­stasis by orchestrating different quality control
Small peptides in mitochondria mechanisms, including protein quality control pathways,
and bacteria with a formylated
amino-terminal Met and,
mitochondrial dynamics, mitophagy, mitochondrial
usually, a hydrophobic amino biogenesis and apoptosis1,73; however, crosstalk between
acid at the carboxy-terminal. mitochondria and the nucleus functions as a parallel
and powerful mechanism that cooperates with these
response mechanisms under normal and stress condi‑
tions. Mitochondrial defects, especially decreases in ATP
production, trigger a pleiotropic retrograde response
that has the main effect of activating alternative path‑
ways of energy generation and restoring mitochondrial
homeostasis. Retrograde signalling to the nucleus and
mitochondrial feedback occur in all eukaryotes; how‑
ever, the complexity of their regulation has evolved in
parallel with the complexity of organisms. Thus, a large
number of components and pathways — which may be
either cell-autonomous or non-cell-autonomous, and
tissue-specific — are associated with mitonuclear com‑
munication in mammals. This plethora of mechanisms
has hampered the study of mitonuclear communica‑
tion regulation and our understanding of the physio­
logical and pathological consequences of the activation
of mitonuclear signalling. Nevertheless, mitonuclear
communication seems to be essential for maintaining
certain physiological functions, such as the immune
response (BOX 1). Future studies using loss‑of‑function
and gain‑of‑function in vivo models, as well as the analy‑
sis of different genetic populations155, including humans,
should help us to understand this complex regulation
in mammals.
The study of mitonuclear communication has
received great interest since the discovery that activa‑
tion of some stress response pathways promotes longev‑
ity. Non-deleterious defects in components of the ETC
activate a paradoxical beneficial response to promote
health and extend lifespan, thereby strengthening and
expanding the mitohormesis theory 156. Additionally,
the fact that this stress response can be regulated in a
non-cell-autonomous manner by mitokines opens up
a new field of investigation around these molecules,
which could function as pharmacological targets for
managing mitochondrial and ageing-related diseases.
Although some hormones and peptides have been
described as potential mitokines, we expect that sev‑
eral other molecules may function as mitokines. As
mitochondria regulate many cellular metabolic path‑
ways, some intermediate metabolites could function
as mediators for both cell-autonomous and non-cell-­
autonomous responses. Long non-coding RNAs,
microRNAs or different kinds of small RNAs could
also mediate mitochondrial-mediated stress responses.
Of note, some long non-coding RNAs, which can be
exported to the cytosol to regulate protein synthesis,
have been identified in mtDNA157. Similarly, mito‑
chondria-derived ­vesicles could be another potential
­communication signal158.
In this Review, we have described the different path‑
ways that are governed by mitonuclear crosstalk under
basal and stress conditions, providing an overview of
these complex networks in various organisms. Although
several evolutionarily conserved mechanisms of stress
response exist, future studies will be required to unravel
the regulation and the molecular mechanisms behind
these stress responses, as well as their physiological and
pathological relevance, which may one day help us to
design potential therapies for pervasive human diseases.

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Acknowledgements
The authors thank all members of the Auwerx laboratory for
their helpful comments on the manuscript and apologize for
the omission of relevant work owing to space constraints.
P.M.Q. is supported by a long-term fellowship from the
European Molecular Biology Organization (EMBO; ALTF
480–2014). J.A. is the Nestlé Chair in Energy Metabolism,
and the Auwerx laboratory is supported by grants from the
École Polytechnique Fédérale de Lausanne, the Swiss
National Science Foundation (31003A‑140780), the AgingX
programme of the Swiss Initiative for Systems Biology
(51RTP0‑151019), the Krebsforschung Schweiz/Swiss Cancer
League (KFS‑3082‑02‑2013) and the US National Institutes
of Health (NIH; R01AG043930).
Competing interest statement
The authors declare no competing interests.

226 | APRIL 2016 | VOLUME 17 www.nature.com/nrm

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