TELOMERES

What are they? Why are they important? Telomere shortening and the end-replication problem Telomerase Telomere hypothesis of aging

Telomeres
Ends of linear chromosomes
Centromere

Telomere

Telomere

Repetitive DNA sequence (TTAGGG in vertebrates) Specialized proteins Form a 'capped' end structure

Telomeres 'cap' chromosome ends

TELOMERE STRUCTURE
5 3

Telomeric t loop
5'
Telomeric proteins: TRF1 TRF2 TIN2 RAP1 TANKS 1,2 POT1 etc

3'
NUCLEAR MATRIX

Why are telomeres important?

Telomeres allow cells to distinguish chromosomes ends from broken DNA

Stop cell cycle! Repair or die!!

Homologous recombination
(error free, but need nearby homologue)

Non-homologous end joining

(any time, but error-prone)

Why are telomeres important?

Prevent chromosome fusions by NHEJ (non-homologous end joining)

NHEJ

FUSION BRIDGE
Mitosis

BREAKAGE Fusion-bridge-breakage cycles

Genomic instability
Cell death OR neoplastic transformation

Telomere also provide a means for "counting" cell division

Proliferative capacity

Finite Replicative Life Span "Mortal" Number of cell divisions

Infinite Replicative Life Span "Immortal"

How do cells "know" how many divisions they have completed??

The End Replication Problem:

Telomeres shorten with each S phase
5' 3' 3' 5'

5' 3' 5'

3' 5'

DNA replication is bidirectional Polymerases move 5' to 3' Requires a labile primer

Ori

Each round of DNA replication leaves 50-200 bp DNA unreplicated at the 3' end

Telomere also provide a means for "counting" cell division: telomeres shorten with each cycle
20

Telomere Length (humans)

Telomeres shorten from 10-15 kb (germ line) to 3-5 kb after 50-60 doublings (average lengths of TRFs) Cellular senescence is triggered when cells acquire one or a few critically short telomeres.

10

Normal Somatic Cells

(Telomerase Negative)
Cellular (Replicative) Senescence

Number of Doublings

How do replicatively immortal cells

avoid complete loss of telomeres

(how do they solve the end-replication problem)?

TELOMERASE:
Key to replicative immortality

Enzyme (reverse transcriptase) with RNA and protein components Adds telomeric repeat DNA directly to 3' overhang (uses its own RNA as a template) Vertebrate repeat DNA on 3' end: TTAGGG Telomerase RNA template: AAUCCC

TELOMERASE:
Key to replicative immortality
+ TELOMERASE

Overcomes telomere shortening and the endreplication problem

Expressed by germ cells, early embryonic cells

Not expressed by most somatic cells (human) May be expressed by some stem cells, but highly controlled Expressed by 80-90% of cancer cells Remaining still need to overcome the end replication problem; do so by recombinational mechanisms -ALT (alternative lengthening of telomeres) mechanisms

Telomere Length and Cell Division Potential

20

Germ Cells (Telomerase Positive)

Telomere Length (humans)

10

Normal Somatic Cells

+ Telomerase

(Telomerase Negative)

Cellular (Replicative) Senescence

Number of Doublings

HOWEVER,
CELLS THAT EXPRESS TELOMERASE

STILL UNDERGO SENESCENCE

(E.G., IN RESPONSE TO DNA DAMAGE, ONCOGENES, ETC.)

Inducers of cellular senescence

Cell proliferation (short telomeres) DNA damage Oncogenes Strong mitogens/ stress

Potential Cancer Causing Events

Telomerase:
Biomedical uses

Expand cells for replacement therapies (burns, joint replacements, etc)

Telomerase inhibitors to selectively kill cancer cells

The telomere hypothesis of aging

Telomeres shorten with each cell division and therefore with age TRUE Short telomeres cause cell senescence and senescent cells may contribute to aging TRUE HYPOTHESIS: Telomere shortening causes aging and telomerase will prevent aging TRUE OR FALSE?

The telomere hypothesis of aging

Telomere length is not related to life span (mice vs human; M musculus vs M spretus)
Telomeres contribute to aging ONLY if senescent cells contribute to aging Telomerase protects against replicative senescence but not senescence induce by other causes

SUMMARY
Telomeres are essential for chromosome stability Telomere shortening occurs owing to the biochemistry of DNA replication Short telomeres cause replicative senescence (other senescence causes are telomere-independent) Telomerase prevents telomere shortening and replicative senescence The telomere hypothesis of aging depends on the cellular senescence hypothesis of aging