CELL CYCLE

Basic of cell cycle is to:

-duplicate accurately the vast of DNA and
- segregates the copied DNA precisely to
two genetically identical daughter cells

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CELL CYCLE
In a somatic cell, cell cycle is composed of;

1. Mitotic phase (M phase)

- mitotic cell division.
2. Gap 1 phase (G1phase)
- cellular differentiation & growth.
3. Synthetic phase (S phase)
- DNA replication
4. Gap 2 phase (G2 phase)
- preparation for cell division.
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Control of
cell cycle

• Cell cycle is driven by cyclin and cyclin

dependent protein kinase (cdk).
• Checkpoint occurs in G1 & G2 phase.
• After ‘M’ phase to at the end of ‘S’ phase
is called ‘interphase’.
• Gene expression is occurred during
interphase.
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Cyclin
&
Cdk

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RESTING CELLS

• In liver
After differentiation, cells enter ‘G0’ phase
(resting phase).
• When condition is favorable or need to
regenerate, enter ‘S’ & ‘M’ phases.
• Muscle is stopped in G0 phase, no
regeneration.

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CELL CYCLE PROGRESSION
(G1 to S phase)

• Cyclin D level rise in G1 phase and

activates CDK4 and CDK6.
• G1 phase has Restriction point, regulated
by retinoblastoma protein (Rb).
• Rb inhibits cell cycle progression by
binding to E2F.
• Cyclin D/CDKs complex phosphorylates
the Rb and E2F is released from Rb.
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CELL CYCLE PROGRESSION

• E2F enhances transcription of gene and

cell cycle progression.
• Cyclin E and A are necessary for the early
S phase.
• In both G1 & G2 phases, cell cycle
progression is also checked by p53
protein.
• Transition from G2 to M phase is driven by
cyclin B. 19
• Cell cycle control at G1 and G2 mechanisms
are working through protein p53

• P53 produces p21 protein which is cyclin-

Cdk inhibitor

• Inhibition halt progression of cell cycle

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Signals

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Biomedical Importance

• Oncogenic viruses can alleviate or

disrupt the restriction point in G1
phase.

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Apotosis

• Program cell death - tightly regulated

cell suicide

• Cells no longer needed or threatening

organisms undergo apotosis

• Mediated by proteolytic enzyme,

caspase, exists in all cells procaspase
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❖ Fragmentation of DNA,

❖shrinkage of ctyoplasm,

❖ membrane changes

❖ No lysis, no damage to surrounding cells.

▪ Initiated by various signals Eg. damaged

DNA

▪ Inhibited by inhibitor proteins and Bcl-2,

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DNA REPLICATION

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 Replication
▪ Replication is the process by which each
strand of the parental DNA duplex is copied
precisely by base pairing with complementary
nucleotides.

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• Replication of DNA genome occurs ‘S’
phase of cell cycle.
• Nuclear DNA is completely replicated
once and only once.
• Replication is - semiconservative.
- semidiscontinuos.
- simultaneous.
- bi-directional.
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Replication is simultaneous and
semidiscontinuous

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DNA Polymerase in eukaryotes

• Polymerase a & d – DNA replication

- Polymerase a – primer formation

- Polymerase d – elongation

• Polymerase b & e – nuclear DNA repair

• Polymerase g – replication of

mitochrondrial DNA 39
DNA Replication in Eukaryotes

• catalyzed by DNA polymerase.

• proceeds in 5’ to 3’ direction.
• needs primer (short strand of RNA).
• before replication, chromosome must
be relax into chromatin.

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 Initiation phase

1. Identification of the origin of

replication (ori)
• chromosomes are converted into
chromatin.
• The nucleosome must be released
into histone and bare DNA strands.

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Initiation phase

• origin of replication (ori) must be

identified on both DNA strands.
• there are multiple sites for replication
origin.

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Initiation phase
2. Unwinding of double stranded DNA
into single stranded DNA
• DNA helicase allows unwinding of
DNA stands.
• single stranded DNA binding
proteins (SSBs) stabilize and
prevent rewinding of DNA strands.

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3. Formation of replication fork
• At the site of unwinding, replication fork is
formed.
• A replication fork consists of four components;
• - Unwinds duplex DNA
- DNA helicase,
- DNA polymerase a (Pol a),
- SSBs.

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• Primer formation; the initiation of
DNA synthesis requires primer, a
short length of RNA.
• Primer is synthesized by DNA Pol a,
which has primase activity.

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dNTP are joined by 3’ 5’ phosphodiester
bond

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 Elongation phase
4. Initiation of DNA synthesis and
elongation
• After forming the primer, DNA Pol
polymerizes the complementary
nucleotides in 5’ to 3’ direction.
• One strand is synthesized continuously;
leading strand.
• The opposite strand is synthesized in
short fragments; lagging strand.
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Elongation phase
5. Formation of replication bubbles
• the multiple replication origins generate
replication bubbles.
• In one replication bubble, DNA synthesis
is bi-directional.
• Supercoiling is occurred due to
unwinding of multiple sites.
• DNA topoisomerase I & II release the
supercoiled DNA.
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Elongation phase

• RNA primers must be removed.

• gaps must be filled with deoxyribonucleotides

by Pol a

• Pol b and Pol e have proofreading activity 5’

to 3’ and 3’ to 5’ exonuclease activities.

• Telomere portions are replicated by

telomerase.
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Telomere replication
Telomerase with bound RNA template

Parental strand
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Termination phase

6. Reconstitution of chromatin
structure
• Chromatin structure must be reformed
after replication.
• One daughter strand and one parent
strand of DNA are rapidly assembled into
nucleosome, and then to chromatin
structure.
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SEMI-CONSERVATIVE

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Post replication modification of DNA

• After DNA is formed it is glycosylated

and methylated by specific enzymes.
• Methylation on adenine residues of
daughter strand protects the host DNA
from destruction of restriction
endonuclease.

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DNA Polymerases in prokaryotes

• DNA polymerase I; participates in

replication and repair.
• DNA polymerase II; participates in
repair.
• DNA polymerase III; participates in
elongation.
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Replication in prokaryotes

• Initiation

- identification of ori C

- binding of dnaB, dnaC and dnaA

proteins.

- dnaB is helicase and needs ATP.

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- unwinding of circular DNA produces
left handed supercoiling.

- DNA gyrase introduces right handed

DNA.

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General scheme of replication

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• Elongation

- primase synthesizes primer formation.

- DNA polymerase III catalyzes elongation of

DNA strands.

- leading strand is continuously grow.

- lagging strand is discontinuously formed.

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• Termination

- primers are removed by 5’ to 3’ exonuclease

activity of DNA pol I.

- gap is filled with dNTP by DNA polymerase I.

- DNA polymerase I is able to do proofreading

because it has 3’ to 5’ exonuclease activity.

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Inhibitors of DNA gyrase

- Quinolones and fluoroquinolone

- Nalidixic acid
- Ciprofloxacin
- Norfloxacin

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 Reverse Transcription

• Formation of DNA from RNA.

• This process is essential for an

organism which carries the genetic
information as RNA

• By reverse transcriptase.

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• Reverse transcriptase can
synthesizes DNA from RNA template.

• It can use RNA or DNA as a template.

• It also has ribonuclease activity

• It is present in retro virus.

• Viral genome can introduce into host

genome.
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Inhibitors of Reverse Transcriptase

• Use in treatment of HIV infection.

• Most of the compounds are nucleotide
analogs.
• Azidothymidine (AZT)
• Dideoxy cystidine (ddC)
• Dideoxy inosine (ddC)

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