Xu Zhaoyang

Department of cell biology

Basic medical college
Zhengzhou University
 Preface
The division cycle of most cells consists of four
coordinated processes:
Cell growth
DNA replication
Distribution of the duplicated chromosomes to
daughter cells
Cell division
 G1 Phase

Metabolically active
Organelle duplication, but no DNA replication
� Duration variable – short in embryonic and
cancer cells
Prepares for S phase
Cells that remain in G1 for a long time = G0
(permanent tissues, such as neural tissue)
 S Phase

� �Committed to cell division once this starts

� DNA and centrosome replication
� Semi-conservative replication of DNA:
two identical daughter genomes
 G2 Phase

�� Growth continues
�� Enzymes and proteins synthesized for
cell division
�� Determining Cell Stage
�� Cells at different stages of the cell cycle can
also be distinguished by their DNA content
 Mitotic (M) Phase

�� mitosis plus cytokinesis

�� Mitosis:
�� Prophase
�� Metaphase
�� Anaphase
�� Telophase
 Regulation of the Cell Cycle

Regulatory signals
Check-points
Cyclins
CDKs
Regulatory signals

Internal signals
DNA synthesis
Nucleotide level

External signals
Growth factors
Hormones
Cell-Cycle Checkpoints
 G1 Checkpoint

• �START in yeast
• �Restriction Point in mammals
• �Main step that commits a cell to division
• �Sensitive to:
• �� Cell size
• �� Availability of nutrients
• �� External growth factors
 G2 checkpoint

��
Error check: DNA replication must be
complete
� Detects unreplicated DNA, holds cell at
G2
�� Detects damaged DNA, arrests cell in G2
until damage repaired
 Spindle Assembly Checkpoint

• Boundary between metaphase and anaphase

• All chromosomes must be properly attached to the
spindle
• MPF causes activation of anaphase promoting
complex pathway that promotes anaphase
 Cyclins
First found in sea urchin;
�Composed of two kinds:
1.Mitotic Cyclins :drive cells into M phase ,
cyclin A,cyclin B,M-cyclin

2.G1 cyclins : drive cells into S phase , cyclin C,

cyclin D��

Level changed correspond to the stage of cell cycle

 Cyclins

Correlated cyclin level Cyclin destroyed every

with cell division cycle time the cell divide
cyclin levels in cell cycle
 Cyclin-dependent
Kinase
Family of serine/threonine protein kinases whick
level remain stable in cell cycle
Play a core function in ensuring correct progression
of the cell cycle.

9 CDKs have been identified, with 5 of these are active

CDK7 acts with cyclinH as a CDK activating kinase (CAK)

 Cyclin-dependent
Kinase
Requires 2 steps to activation:
Partial activation occurred after specific cyclin
binding
complete activation requires phosphorylation by a
CDK-activating kinase (CAK)

The result of activation :Induce downstream

processes by phosphorylating selected proteins
 Progression regulatory
of cell cyclye

• 1.Cyclin binding
• 2.CDk phosphorylations state
• 3.Cdk inhibitors
• 4.Controlled proteolysis
• 5.Sub cellular localization
 Cyclin binding

• The binding causes a major change in the

conformation of the catalytic subunit and
the movement of flexible loop of the CDk
polypeptide chain away from the opening
to the enzyme’s active site, allowing the
CDk to phosphorylate its protein
substrates.
 CDk phosphorylations state

• CDk + cylclin + phosphorylation =

active

• The cyclin-CDk complex to be active it

has to be phophorylated at one or
more sites by a specific protein
kinase, and de phosphorylated at
other sites by a specific protein
phosphatase.
Mechanisms of Cdk
regulation
 CDk inibitor

INK4 proteins
Specifically inhibit the catalytic subunits of CDK4 and
CDK6
p16INK4a , p15INK4b , p18INK4c , and p19INK4d
Cip/Kip family
Inhibitors of cyclin E- and A-dependent CDK2
Positive regulators of cyclin D-dependent kinases
p21Cip1 , p27Kip1 , and p57Kip2 , all of which contain
characteristic motifs within their amino-terminal moieties
that enable them to bind both to cyclin and CDK subunits.
 Controlled proteolysis

• SCF complex
• Active from late G1 to early mitosis
• Mediates the destruction of G1 cyclins ,
Cdk inhibitors, et al.
• APC complex
• Degrade a nubmer of key mitotic
proteins, including the mitotic cyclins.
Destrction of them allows a cell to exit
mitosis and enter a new cell cycle.
Sub cellular localization
Confocal immuno-fluorescent images of
asynchronous HeLa cells, labeled with Cyclin
B1 (V152) Mouse mAb(green) showing
cytoplasmic localization of Cyclin B1 in pre-
mitotic cells, translocation into the nucleus in
early mitosis and degradation of Cyclin B1 in
anaphase.
Its nuclear accumulation is facilitated by
phosphorylation of a cluster of serine
residues that resides in its nuclear export
signal, presumably blocks subsequent
export of the cyclin back to the cytoplasm
and initiates cell division.
 Cyclin-CDKs in different
stages in the mammalian
cell cycle
Essential for
the entry to G1 Induced by cyclinD,
Regulates progression
from G1 to S

Promotes G2 to M
entry with CDK1

Mitosis
regulated by CDK7 binds with
CDK1 binding Cyclin H
throughout the cell
cycle
 Cell cycle and tumor

Oncogene

Tumor suppressor gene

Tumor treatment
 The production of
oncogene
1.Growth factors
2.Growth factor receptors
3.Protein kinases or
proteins that activate
protein kinases
4.Proteins that control
cell cycle
 Tumor Suppressor Gene
Tumor Function Chromosomal Tumor Types
Suppressor Location Observed
Gene

P53 cell cycle regulation, 17p13.1 sarcomas, leukemia,

apoptosis breast cancer

RB1 cell cycle regulation 13q14.1-q14.2 retinoblastoma

p16INK4a cell-cycle regulation 9p21 melanoma,

protein=CDK pancreatic cancer
I2A
 p53

Tumor Suppressor Gene

50% of all cancers related to mutations of p53
Called “guardian of the genome”

�DNA damage increased levels of p53

�p53 is a transcription factor activates gene
coding for p21(cdk inhibitor that arrests cells in G1
until damage repaired),If DNA repair not
successful, p53 induces apoptosis.
��
�
Inhibitor of Cell Cycle Progression

Induction of p21 by DNA damage

 G0
EGFR
PDGFR
Cyclin D
CDK4,6
G1a
M
p16, p15 Cyclin E
G1b CDK2

Cyclin A,B p53 p21

CDK2
p27

pRB G1c
G2 E2F
p21
p27 G1d
S
Cyclin
CDK2A Cyclin E
CDK2
 Rb protein

�� Rb gene: expresses Rb protein, acts as a

G1 brake
Key substrate of Cdk-cyclin D complex
� Binds to transcription factor E2F
� Growth factors activated Cdk-Cyclin
� Phosphorylates Rb Allows transcription to
proceed
��
The roll of RB
 The treatment of tumor
Antimutagenesis mechanisms
Antiproliferative Mechanisms
Induce apoptosis
Inhibit angiogenesis
Inhibit polyamine synthesis
Inhibit oncogene activity
Induce terminal differentiation
Inhibit DNA synthesis
Balance DNA methylation
Modulate growth hormone activity
Agents
Retinoids
Retinoids, tamoxifen, NSAI
Difluoromethylornithine
Perillyl alcohol, FTI
Retinoids, Vitamin D
Oltipraz, genistein
Folic acid, budesonid
Tamoxifen, soy isoflavones