Several studies indicate that ROS promotes telomere shortening and cellular senescence.

The
telomere shortening rate and the onset of cellular senescence of MRC5, WI-38 or IMR-90
human fibroblasts correlated with oxygen concentration to which the cells were exposed to in
tissue culture chambers (Ahmed et al., 2008; Forsyth et al., 2003; Passos et al., 2007). Since
non-transformed human fibroblasts do not express telomerase the results suggest that
oxidative damage enhanced the end replication problem interfering with the maintenance of
the distal ends of the telomeres. Specifically, one could speculate that ROS induces by
unknown mechanisms an increased length of the telomeric 3’ overhang or it might trigger
premature abortion of replication before reaching the end of the DNA template. Enhanced
susceptibility of the distal telomeric template strand to oxidative damage should cause the
latter phenomenon as oxidative base damage or base loss can block polymerases and can
cause replication fork stalling that upon collapse leads to double-strand breaks. Instead,
oxidative damage at random positions within telomeres may result in stochastic and more
irregular telomere shortening events (von Zglinicki et al., 2000). In addition, breakage of the
DNA backbone can also be provoked by ROS (Aeby et al., 2016) leading to telomere
truncation when not repaired before DNA replication. Acute damage at telomeres by ROS
and cellular senescence was also provoked experimentally in a study in which KillerRed, a
fluorescent protein generating ROS upon illumination with visible light was recruited to
telomeres via gene fusion with TRF1 (Sun et al., 2015).
In addition to interfering with semiconservative replication of telomeric DNA,
ROS-damaged DNA or oxidized dNTP precursors for telomere synthesis interfere
with telomerase activity (Figure 2). 8-oxodGTP is used by human telomerase as a
substrate but upon incorporation it functions as efficient chain terminator (Aeby
et al., 2016; Fouquerel et al., 2016). Similarly, an oligonucleotide DNA substrate
ending in TTA-8oxoG-3’ instead of TTAG-3’ could not be extended by purified
human telomerase (Aeby et al., 2016). Then again, a DNA substrate ending with
TTAGG-8oxoG-3’ was extended efficiently. Therefore, depending on the position
within the DNA substrate, the outcome is different. For the inactive substrate, the
oxidized nucleotide corresponds to the position in which the substrate reaches
the end of the RNA template and must be realigned at the other side of the
template for further extension suggesting that this step is perturbed by 8oxoG
(Figure 2B). Therefore, these studies provide a molecular explanation for how
ROS interferes with telomere extension by telomerase in human cells (Forsyth et
al., 2003).
Under certain circumstances oxidative damage of telomeric DNA might also
promote telomere elongation by telomerase. ROS-damage of telomeric DNA will
disrupt binding by TRF1 and TRF2 (Opresko et al., 2005). Furthermore, because
8oxoG frequently mispairs with adenine during replication the mutated
telomeric DNA sequence will interfere with binding of shelterin components and
consequently telomere function if prevalent. Since experimental depletion of the
shelterin components TRF1, TRF2 or POT1 increases telomere length (Ancelin et
al., 2002; Loayza and De Lange, 2003; Smogorzewska et al., 2000; van Steensel
and de Lange, 1997), a reduction of shelterin binding sites at telomeres is
predicted to increase telomere extension efficiency by telomerase. In addition, 8-
oxoG and thymine glycol (Tg)-containing telomeric DNA substrates disrupt or
alter G-quadruplex structures (Fouquerel et al., 2016; Lee et al., 2017), which are
inhibitory to telomerase. Thus, the modulation of telomeric DNA secondary
structures through oxidation of DNA bases may also positively influence
telomerase, as long as oxidative damage does not interfere with telomerase at
the very 3’end. However, the current analyses in cells and animals suggest that in
vivo the negative effects of ROS on telomere length prevail.
The biology of genome stability and telomeres includes many interactions between these two
factors. Short and dysfunctional telomeres are the initiating point for cell senescence, cell
death, and DNA
instability (18). The effect of aging on the level of DNA damage in
human cells has been assessed in many studies (19,20). Over time,
genome damage accumulates probably due to lower DNA repair
capacity, lower chromosome segregation, and cell cycle checkpoint efficacy (21). Unhealthy
factors, which include endogenous
genotoxins, inadequate nutrition, and other unfavourable lifestyle
factors, are also responsible for both an increase in baseline genome damage and accelerated
telomere shortening. Most of these
factors have an indirect mechanism of action through free radical
formation and oxidative stress. This points to a conclusion that
since reactive oxygen species (ROS) are predominantly responsible
for age-related damage at the cellular and tissue levels (22,23), a
reduction in calorie intake would lead to a reduction in energy
metabolism and ROS production. Consequently, this would result
in diminished cell and tissue damage. Indeed, ROS strongly influences replicative senescence
and aging through accelerated telomere shortening caused primarily by accumulation of nicks
in the
G-rich strand (24).
Under physiological conditions, mitochondria are the main
source of ROS and it has been demonstrated that senescence of
human cells is related to dysfunctional mitochondria and shorter
telomeres (25) (Figure 1). Overproduction of ROS results in oxidative stress which can lead
to oxidation of biomolecules including
lipids, proteins, and DNA over time. Such loss of DNA structure or
its information content typically cannot be replaced (26). Obviously,
there is a valid reason why chronic oxidative stress is perceived as
one of the leading causes of many metabolic and neurodegenerative
diseases and chronic inflammation. Inflammation causes accelerated
white blood cell turnover and additional telomere attrition resulting
in accelerated aging (27,28).
Furthermore, mitochondrial and telomere mechanisms of aging
have been united and a direct molecular connection between telomere
attrition and mitochondrial dysfunction has been established (29). It
has been shown that p53 activation induced by telomere shortening,
in addition to halting the cell cycle, downregulates the expression of
PGC1α and β which leads to reduction of mitochondrial biogenesis
and function. Dysfunctional mitochondria demonstrate metabolic
changes, such that they exhibit defective ATP generation, increased
ROS production, and decreased levels of ROS-detoxifying enzymes
which cause additional damage both to mitochondrial and genomic
DNA, including telomeres (29) (Figure 1)
Besides p53 protein, sirtuin family of NAD-dependent protein
deacetylases and ADP-ribosyltransferases plays an important role
in control of mitochondrial function. SIRT1 modulates mitochondrial biogenesis through
deacetylation and activation of PGC-1α
while SIRT3 deacetylates mitochondrial enzymes involved in energy metabolism linked to
some of the beneficial effects of calorie restriction (1)
Also, mitochondria can contribute to aging independently of
ROS, through numerous types of mitochondrial dysfunctions like
impaired mitochondrial biogenesis, decreased mitophagy and fission
fusion ratios, mutations and deletions in mtDNA or perturbations
on electron transport chain. All this can cause reduced efficiency of
mitochondrial bioenergetics (1).
Nevertheless, it should be emphasized that ROS are normal
intermediaries of biochemical reactions which serve as important
physiological regulators of intracellular signaling pathways (30).
Depending on the circumstances, they can be either harmful or beneficial to living systems
(31). Beneficial effects of ROS occur at low/
moderate concentrations and involve physiological roles in cellular
responses to anoxia, defense against infectious agents, or induction
of a mitogenic response (30). In contrast, a significant increase in
oxidative stress induces premature senescence in many human cell
types (32). Thus, the delicate balance between the beneficial and harmful effects of free
radicals is a very important aspect of human
aging (33) as proposed in the gradual ROS response hypothesis (23).