L24: Telomeres, Telomerase, and Aging

Learning objectives:
• Understand the basics of DNA replication
• Understand the end-replication problem
• Know that telomeres are heterochromatic repetitive
DNA sequences
• Learn that telomerase is a ribonucleoprotein that
elongates chromosome ends
• Understand the concept of cell senescence
• Realize the connection between telomere attrition,
stem cell senescence, and organismal aging
• Be exposed to the nine proposed hallmarks of aging
• Realize the vast body of research leading to proposed
aging remedies
Essential Cell Biology, Fifth Edition
Copyright © 2019 W. W. Norton & Company
• BIOMG1350 PRELIM 4: December 4th, 2022 (L19-24; S9-11)
• check your room/seat assignment by Saturday
• practice prelim uploaded
• review sessions with TAs 1-4PM this Sat/Sun in G01 Biotech

• BIOMG 1350 FINAL EXAM: Saturday, Dec 9th at 7:00 PM in Barton Hall. BTN100CENT, BTN100EAST
• We made previous prelims available for practice in Canvas/Exams
• Martin Graef office hours Wed Dec 6th at 4pm zoom;
• Doina Tumbar office hours Thu Dec 7th at 7pm on zoom
• Review sessions with TAs 2-4PM December 7th and 8th - Thursday and Friday - Biotech G01
• Watch for final instructions announcement next week
• It is your responsibility to read and know the rules, including academic integrity issues

• BIOMG 1350 MAKE-UP FINAL EXAM REQUEST: DEADLINE Nov 29th

• send request to biomg1350@cornell.edu with
 a reason for wanting a make-up exam
 a list of ALL your final exams with their course number, date and time
 conflicts will be scheduled between Dec 13th - 17th, date TBD
Macromolecule Macromolecules Cellular Structures Developmental Biology Stem Cell Biology
Building Blocks & Synthesis & Processes & Embryogenesis & Homeostasis

The Biology of Aging

 Adult Stem cells

Embryonic stem cells

Stem Cell Lecture’s
Topics Reprogramming

Applications
 Basic Science Applications of ESCs:
The Nobel Prize in Physiology/Medicine 2007 for introducing specific gene
modifications in mice via embryonic stem cells

Gene targeting
to introduce mutations
(e.g. CRISPR/Cas9)

Mutant ES cells

Breeding to transmit mutation

to next generation of mice

Blastocyst develops in
Clump of mutant foster mother into a
ES cells injected in chimeric mouse; the ES cells
blastocyst contribute to all tissues
including germ line
Figure 22-40 MBC6e
Example of gene knockout experiment:
leptin knockout mice are obese

* KO experiment helped show that leptin is a hormone

which regulates body fat as well as the sensation of
hunger, and its loss results in obesity, susceptibility to
diabetes, and other complications.
Reprogrammed skin cells combined with gene therapy
can treat sickle cell anemia in a mouse model

Infect with viruses

expressing the three
transcription regulators

DevBio11 5-24
 Organoids: study human mini-organs in a dish using iPSCs

Organogenesis

DevBio11 5-25
Adult hematopoietic stem cells are frequently used in clinics

Find immune-compatible Inject the cells into the

Use strong chemotherapy or donor and collect stem cells bloodstream of the patient;
radiation to kill the cancer to transplant. (In some cases, stem cells will find their
cells, but this treatment also patient can donate marrow way to the marrow and
kills hemopoietic stem cells. prior to treatment.) repopulate the blood.
Macromolecule Macromolecules Cellular Structures Developmental Biology Stem Cell Biology
Building Blocks & Synthesis & Processes & Embryogenesis & Homeostasis

The Biology of Aging

 How long can we live?

The oldest age to which any person on record has lived was a French woman,
Madam Jeanne Calment, who was born in the small town of Arles on February 21,
1875 and died August 4, 1997 at age 122 years, 164 days
What are the causes and how can we prevent aging?

An important contributor to aging is the inability to fully

replicate and hence maintain telomere length in tissue stem
cells (along with several other causes).
 Telomeres are heterochromatic regions devoid of genes
and made of repetitive DNA sequences

Telomere length shrinks with age due to incomplete replication at each cell division ->AGING?
 DNA acts as a template for its own replication

• Base-Pairing Enables DNA Replication

• A free 3’ hydroxy (OH) is needed to add a new nucleotide
• DNA polymerase synthesizes complementary new strands in the 5’->3’ direction
Essential Cell Biology, Fifth Edition
Copyright © 2019 W. W. Norton & Company
 Major steps for DNA synthesis (e.g., replication)

DNA Synthesis Begins at Replication Origins

Replication bubble with right& left ‘replication

forks’
RNA primers

3’HO- 5’ Enzyme primase synthesizes RNA primers in 5’-3’

5’ -OH3’
direction providing a free 3’ hydroxy group
Templates ready for DNA synthesis
by DNA polymerase alpha

Essential Cell Biology, Fifth Edition

Copyright © 2019 W. W. Norton & Company
 DNA-polymerase uses the 3’hydroxy on primer to start
lagging strand DNA synthesis in 5’-3’ direction
5’
Replication starts immediately and is
continuous on the leading strand 5’ -OH3’
3’
leading strand Primase adds primers on lagging strand in 5’-3’ directio
and DNA polymerase begins DNA synthesis

Replication is delayed and fragmented on the

lagging strand
Okazaki fragments

The end replication problem

RNA Primers Removed by Nuclease
• The leading strand is synthesized in its entirety. and Replace with DNA by DNA-polymerase
• The lagging strand cannot be replicated to the end Gaps sealed by ligase
• W/O a special mechanism ~100bp DNA would be lost
at each cell division -> 70-100 divisions telomeres
would be gone
 Telomerase: a ribonucleoprotein that replicates the ends of
eukaryotic chromosomes

RNA (TERC or TR):

Used as template

(in San Diego,

California, USA)

Nobel Prize for Medicine in 2009 protein (TERT): reverse transcriptase

synthesizes DNA from an RNA templat
Telomeres and telomerase prevent linear eukaryotic chromosomes from shortening with each cell division

Essential Cell Biology, Fifth Edition

Copyright © 2019 W. W. Norton & Company
 DNA REPLICATION: SUMMARY
• Base-Pairing Enables DNA Replication
• DNA Synthesis Begins at Replication Origins
• Two Replication Forks Form at Each Replication Origin
• DNA Polymerase Synthesizes DNA Using a Parental Strand as a Template
• The Replication Fork has a leading and a lagging DNA strand
• Short Lengths of RNA Act as Primers for DNA Synthesis
• DNA synthesis on the lagging strand occurs in fragments (Okazaki)
• Nucleases and ligases cooperate with primase and DNA polymerase to synthesize
DNA on lagging strand
• The end-replication problem = incomplete replication on the lagging strand
• Telomeres are heterochromatic regions made of repetitive sequences
• Telomerase extends the chromosome ends from its RNA template
 Figure 2

Telomere attrition results in

permanent cell cycle arrest
(i.e., cell senescence)

• Cells w/o telomerase reach critically short

telomeres and stop their proliferation
• This is called the “Hayflick limit” and results in cell
senescence, a form of cellular ‘aging’
• Senescent cells do not die but they permanently
withdraw from the cell cycle
• Senescence is a form of tumor-suppression
• Restoring telomerase activity in these cells restores
their proliferative potential
• Cells with high telomerase activity are immortal

Current Biology 1998 8R178-R181DOI: (10.1016/S0960-9822(98)70105-8)

 Telomere length and telomerase activity varies in different
cell types

T is s u
e

Pr ffer
og en
Di

en tia

Telomerase activity
ito te
ra dc
n d el l
s

https://www.spandidos-publications.com/ijo/43/5/1351
 Senescence of tissue stem cells may contribute to aging
TERC Knockout Mice:
• Early generation : no phenotype due to mice having very long telomeres
• Late generations: phenotypes show defects in tissue homeostasis
- Mice viable for several generations (up to 5)
- Male and female infertility increases with generation- germline SCs apoptosis
- Diskeratosys congenita—like phenotypes (a disease with some characteristics of
premature aging) due to limited tissue SC proliferation
• Defective closure of neural tube (NSC)
• Small size/atrophy of the intestine (ISC)
• Abnormal skin pigmentation (MSC)
• Defective skin/nail/hair regeneration (HFSC)
• Bone marrow failure (HSC)
• Death
TERT Over-expression Mice: Live longer and have elevated SC activity (but have
higher incidence of cancer)
Telomerase and telomere length represent only one factor
contributing to organismal aging

A few examples of candidate aging factors are:

• Epigenetic changes and spurious transcription
• Accumulations of mutations due to replication errors and other
challenges
• Changes in the ECM or cellular composition of the stem cell niche
• Altered signals that perturb cell-cell communication
• Endocrine signals (young vs old blood)
Circulatory factors contribute to tissue stem cell decline and organismal aging
‘Heterochronic parabiosis’ = joining blood circulation of old and young mice
Old Young Old Young

Exposing aged mice to a more

youthful systemic environment
promoted stem cell activity and
tissue regeneration/activity

Exposing young mice to an

aged systemic environment
induced impaired stem cell
activity and tissue decline
Endocrine signaling
 Nine proposed hallmarks of organismal aging

Cell. 2013 June 6; 153(6): 1194–1217. doi:10.1016/j.cell.2013.05.0

 Many Thanks to the Fall 2023 Teaching Team!!
Instructors Teaching Assistants Active Learning

Prof. Doina Tumbar Heather Phillips Simian Cai Jingyi Wu Elena Peot
Dr. Cora Demler
BIOMG1035

Prof. Martin Graef Julia Zhu Alexandre Miaule Rachel Bradley

Cristina DeOliveira Dr. Elizabeth Ogata
Active Learning Sections
Thank you!
Best of luck