Title

Shortening of telomeres and other factors related

to the aging process



















2


Table of Contents


Introduction ............................................................................................................................... 3
1. Genetic Theories .................................................................................................................... 4
1.1 DNA Damage Theory ....................................................................................................... 4
1.2 Telomeres ........................................................................................................................ 5
1.2.1 The Hayflick limit ...................................................................................................... 5
1.3 Telomere shortening and replicative senescence ........................................................... 5
1.4 Telomerase ...................................................................................................................... 6
1.4.1 Activating of telomerase .......................................................................................... 6
1.4.2 TERT .......................................................................................................................... 7
1.4.3 TRF2 .......................................................................................................................... 7
1.5. Aging Genes .................................................................................................................... 7
2. Biochemical theories ............................................................................................................. 8
2.1 Reactive Oxygen Species (ROS) ....................................................................................... 8
2.2 Mitochondria .................................................................................................................. 9
3. Physiological theories .......................................................................................................... 10
3.1 Hormones ...................................................................................................................... 10
3.2 Proteins.......................................................................................................................... 10
Conclusion ............................................................................................................................... 11
References ............................................................................................................................... 12








3

Introduction

Aging is a universal and multifactorial bilogical process. The array f changes in
an organism resulting t the aging phenotype is enormous and complex. Thus, several
molecular mechanisms f aging have been proposed and generated a great number f
theories. This reprt will analyze the most prevalent f these theories.
The first section focuses n Genetic theories that deal with accumulation of errors
in the DNA, programmed senescence and telomeres, and aging genes. The second
section discusses the Bichemical theories that are concerned with energy
metabolism, generation f free radicals and mitochondria. And the final section,
examines Physiological theories that deal with the rle of hormones in regulating the
rate of cellular senescence.
As the elderly population rises, the importance of elucidating these mechanisms is
becoming more apparent. Aging research will prvide unique insights into aging
processes and enable new treatments that will help people live longer and healthier
lives.

















4

1. Genetic Theories

1.1 DNA Damage Theory

Most biological theories of aging assume that accumulation of DNA
damage/mutations play a crucial role in aging. DNA Damage theory of aging is
concerned that aging results from the accumulated point mutations and errors in DNA
sequences that accumulate with age over the course of an organisms lifespam. DNA
damage is widely thought to result of oxidative stress, exposure to oxygen radicals, as
a consequence of cellular metabolic functions or by radiation ( Moskalev et al. 2013).
Cells have elaborate repair systems and strategies that detect and repair DNA
damage, but as we age, error correcting mechanisms become less efficient. The
resulting alteration in DNA, including loss of gene function, is thought to be a major
cause of senescence in cells (Clark 1999: 38-39).
Every day, in a typical human cell, thousands of nucleotides are being damaged by
spontaneous chemical events, environmental pollutants, and radiation. In many cases,
it takes only a single defective nucleotide within the coding region of a gene to
produce an inactive, mutant protein.. Some spontaneous mutations occur after
replication as a result of deamination and depurination, if the damage left unpaired the
mutated genes will be passed on to all daughter cells ( Panno 2010:177).
It is concluded that the net result in terms of damage over time reflect a balance
between the rate of production of oxygen radicals and the effectiveness of prevention
or repair of the cellular damage they cause particularly to DNA. Failure of cellular
DNA repair mechanisms is assumed to be a major factor in cellular aging
(Uittenboogaard, et al. 2013).






5

1.2 Telomeres

The telomere theory of aging suggests that the gradual shortening of chromosomes
is responsible for the cellular damage and dysfunction. Telomeres are repetitive DNA
sequences located at the ends of chromosomes and become shorter with each mitotic
cell division. Telomeres do not contain genes but are made of short stretches of DNA
sequences that are repeated over and over. In human telomeres, for example, the base
sequence TTAGGG is repeated thousands of times (Freeman, Quillin and Allison
2013: 295-296).
Telomeres are essential for chromosome replication and genome integrity because
they protect against instability promoting events as degradation of the terminal
regions of chromosomes, fusion of a telomere either with another telomere of with a
broken DNA end, or inappropriate recombination (Granotier and Boussin 2011).

1.2.1 The Hayflick limit

The Hayflick limit (or Mortality Stage 1), is the point in which cells have lost vital
DNA code and cannot be reproduced further. The Hayflick limit describes the
maximum number of times a cell population can divide, which is related to telomere
length. Leonard Hayflick is the scientist who was the first to notice that normal cells
cannot divide indefinitely in culture. It is observed that the length of telomeres chains
becomes shorter as an individual grow older. Eventually the telomeres become so
short that cell replication produces lethal errors or missing pieces in the DNA
sequence, ending the cell; ability to replace itself (Saltsman 2005: 64-65).

1.3 Telomere shortening and replicative senescence

The replication of telomeres by conventional mechanisms is inevitably incomplete,
leading to telomere shortening at each round of cell division (Granotier et al. 2011).
This occurs because the enzyme that duplicates the DNA (DNA polymerase) has to
have some portion of the chromosome out ahead of it (Panno 2011: 39). Telomeres
6

shorten as we get older, they start out 15,000 base pairs long and gradually get down
to roughly 3-5,000 base pairs (Lewis 2009).
For instance, the telomeres in human fibroblast are long enough to permit about 50
rounds of DNA replication that means that the cell can divide about 50 times in
culture (Saltsman, 2005:64-65). Cancer cells, on the other hand, can divide
indefinitely, and from them scientists isolated an enzyme called telomerase. If the
telomerase gene is added to normal fibroblasts, they are no longer bound by the
Hayflick limit and can divide indefinitely like an immortal cancer cell (Lodish 2000).

1.4 Telomerase

Telomerase is an enzyme that contains a protein reverse transcriptase (TERT) and a
template-containing RNA component (TR), found in germ and cancer cells, acts to
restore the shortened telomeres. Telomerase is required for the maintenance of
telomere repeats in most eukaryotic organisms (Autexier and Lue 2006).
Telomerase synthesizing multiple tandem repeats of DNA (called telomeric DNA)
encoded by its RNA template which dictates the synthesis of the G-rich strand of
telomere terminal repeats (Freeman and Quillin 2011: 295-296). Telomerase also
compensates for the erosion of DNA ends during replication and provides the docking
sites for telomeric proteins that bind specifically to the ends of chromosome to
distinguish them from broken DNA ends (Nandakumar and Cech 2013).

1.4.1 Activating of telomerase

In 2009 Nobel Prize in Physiology or medicine awarded to Elizabeth H. Blackburn,
Carol W. Greider and Jack W. Szostak for the discovery of how the ends of
eukaryotic chromosomes, telomeres, are maintained by a specialized reverse
transcriptase, telomerase. The activating of telomerase extend the life of cells beyond
the hayflick Limit. However in mammals, telomerase is active in the germ line and in
stem cells, but its expression in somatic cells may lead to or predispose to cancer
(Blagosklonny et al. 2010).


7

1.4.2 TERT

In 2008 scientists to circumvent this problem they constitutively expressed
telomerase reverse transcriptase (TERT), one of the components of telomerase, in
mice engineered to be cancer resistant by means of enhanced expression of the tumor
suppressors p53, p16, and p19 ARF. TERT over-expression produced a systemic
delay in aging accompanied by extension of the median life span. It is concluded that
constitutive expression of TERT provides antiaging activity in the context of a
mammalian organism (Loba et al. 2008).

1.4.3 TRF2

Recent studies have suggested a role for shelterin component TRF2 in overhang
generation at leading end telomeres. T loops protect the 3 ends of chromosomes,
making them inaccessible to nuclease and enzymes that repeat double strand breaks.
TRF2 contributes to the generation/maintenance of this overhang (Nelson and Cox
2008: 1056-57). When TRF2 is deleted the telomeric overhang signal is rapidly lost.
These observations led to the proposal that TRF2 recruits or activates a nuclease that
generates the 3 overhang at leading - end telomeres (Wu et al. 2010).

1.5 Aging Genes

Research suggests a telomerase based genetic clock may be located on the first or
fourth of the 23 homologous chromosome pairs found in humans. Other genes that
appear to affect the aging process include the Sirtuin gene family (which code for Sir2
proteins), RAS genes and GATA transcription factors (Magalhes et al. 2008).




8

2. Biochemical theories

2.1 Reactive Oxygen Species (ROS)

According to this theory, ROS are toxic byproducts of aerobic metabolism that
induce oxidative damage to various cellular macromolecules due to their high
chemical reactivity (Bratic and Larsson 2013).
Oxygen is a toxic exhaust produced by mitochondria during the metabolic process
of oxidative phosphorylation. This process produces ATP, the energy that cells need
to survive but it also generates and highly reactive compounds known as free radicals
or reactive oxygen species (ROS). Free radicals have an odd number of electrons and
are therefore highly reactive. The oxygen free radical can remove an electron from
virtually any molecule in the cell, including DNA, RNA, proteins and lipids in the cell
membrane. That means, that oxygen radicals can attack and deform protein
molecules, disrupting structural complexes and inhibiting important enzymatic
functions. Oxygen radicals also attack the individual nucleotide bases that make up
both nuclear and mitochondrial DNA (Abheri 2010).
Normally, the body can handle free radicals, but if antioxidants are unavailable, or if
the free radical production becomes excessive, damage can occur. For instance the
enzyme superoxide dismutase (SOD) neutralizes oxygen free radicals as they are
produced but not all of them. As a result, the amount of free radicals increases
gradually with age as the efficiency of SOD is gradually reduced. Many scientists
believe that free radicals are directly responsible for cellular senescence contributing
significantly to the aging process (Panno 2011: 42-43).








9

2.2 Mitochondria

The mitochondrial theory of aging proposes that mitochondria are the source of the
majority of reactive oxygen species (ROS) produced by the cell. As mitochondria
supply about 90 percent of the metabolic energy used in the form of ATP they
become the prime targets for DNA damage from oxygen radicals (Panno, 2011: 38).
Mitochondria continuously generate potentially damaging reactive oxygen species
(ROS) during oxidative phosporylation and these ROS are continuously detoxified by
cellular antioxidants defense enzymes. However, when ROS are excessive the
resulting cellular damage can drive cellular senescence (Velarde et al. 2012).
Mitochondrial function has long been recognized to decline during aging concomitant
with the appearance of mitochondrial morphological alterations. The gradual
oxidative degradation of mitochondria have thus been proposed to play a critical role
in the aging process (Berdanier 2006).
Recent experimental work has provided clear evidence that somatic mtDNA
mutations can cause a variety of ageing phenotypes and create RC (Respiratory
Chain) dysfunction. (Terzioglu and Larsson 2006). Age related cumulative damage to
the mt DNA causes a concomitant decrease in mitochondrial function and perhaps a
further increase in the mitochondrial production of damaging free radicals, leading to
a vicious cycle of an escalating ROS production and mtDNA damage. In fact, it has
been estimated that the level of oxydatively oxidized bases in mtDNA is 10 to 20 fold
higher than that in nuclear DNA (Tosato et al.,2007). Mitochondrial components,
including the mitochondrial genome, as well as the membranes and proteins central to
mitochondrial function accumulate damage with age. The reduced mitochondrial
bioenergetic capability, as well as the increasing oxidative damage to cellular
components, is hypothesized to contribute to the pathophysiology of aging (Bratic and
Larsson 2013).
.. As a conclusion, it is clear that mitochondria play a critical role in aging.
Mitochondrial structure and function decline with age, shown a causative link
between mitochondrial dysfunction and major phenotypes associated with aging
(Masoro 2006: 124-125).


10

3. Physiological theories


3.1 Hormones

Through adult life all physiological functions gradually decline. Several hormones
gradually decrease with age. The age-related change in hormone concentrations has
been hypothesized to play a critical role in the loss of muscle mass and muscle
strength with aging (Visser et al. 2003). Hormones such as thyroid hormones,
estrogens, and glucocorticoids play important roles in cell growth and differentiation.
Furthermore are also important regulators of mitochondrial biogenesis. Age-related
decline in RC function may at least partly be caused by a decline in hormonal levels.
(Bratic and Larsson 2013).
Many therapies that include GH, estrogen or testosterone supplements have
alleviated some of the symptoms of old age, but they have not been able to reverse the
aging process. (Lamberts 1997).

3.2 Proteins
As we age, there is a diminished capacity for cellular protein synthesis. One typical
example is collagen, a fibrous protein that is the most abundant protein in mammals,
constituting 25% of the total protein mass. It is secreted by connective tissue cells, as
well as by a variety of other cell types (Alberts et al. 2002). Collagen is a major
component of connective tissue and accounts for more than 60 percent of the protein
in bone and cartilage. Collagen also provides 50 to 90 percent of the dry weight of the
skin, ligaments and tendons (Lewis 2009).
Because collagen is so widespread in the body, the general degradation of this
extracellular matrix is thought by many scientists to be a major impediment to
efficient functioning of many organs, and a major contributor to overall senescence.
(Panno 2011: 39).




11

Conclusion

Aging is a complex phenomenon for which there is not a universally accepted
explanation. Aging is thought to be a degenerative process caused by accumulated
damage that leads to cellular dysfunction. This cumulative rate of damage increases
in speed and intensity with advancing age.
In future, by studying aging mosaics, scientists hope to gain a deeper insight into the
aging process at the molecular level. The results of these studies are expected to
revolutionize our understanding of the aging process, giving vital information for the
development of therapies designed to slow or reverse the aging process.



















12

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