The Cell
Cycle
Indwiani Astuti
Fac. of Medicine Gadjah Mada University
Yogyakarta
Introduction
• DNA duplication • The cell cycle divided
• cell division into four phases
• the phases have been – the first gap phase (G1)
known 40 years but – DNA replication (S)
last 3-4 years - the – the second gap phase
(G2) -- interphase
genes involved in
– mitosis (M)
control of cycling
process
Correct spindle
Initiation of formation & The length of the
mitosis metaphase plate cell cycle
around 16-24 h.
G2 DNA G2
damage G1
checkpoint
START or
restriction point
DNA synthesis
completion
S
G1/S DNA damage
checkpoint
The cell cycle showing checkpoints at which
DNA is monitored before the next stage of
the cycle is entered
• G1 preparing to
synthesize DNA,
biosynthesis of RNA
& proteins
• S phase, DNA is
replicated & histones
are synthesized. At the
end of S phase DNA
doubled &
Chromosomes
replicated
• G2, cells are preparing for
cell division, replicated
DNA complexes with
proteins, biosynthesis
continues
• nucleus & cytoplasm divide
during mitosis
• two daughter cells are
produced - can begin
interphase of new cell cycle
• cells can also enter a resting
phase (G0)
• External stimuli: growth • Number of recognized
factors (to enter into checkpoints:
G1): GF -induced • START or restriction
signals and GF inhibitors (R) point (late in G1)
(TGF beta) • during S phase
• The progression of cycle • at the G2 to M
from one stage to next is transition to monitor
the completion of
controlled by cyclin- DNA synthesis
dependent kinases • prevent chromosome
(CDKs) & segregation if not
CDKInhibitors intact
• -> Signal transduction • the degradation of
various cyclin
Cell Cycle
Control Genes
Introduction
Two further groups of genes play a major role in
the development of cancer :
1. genes those intimately involved in the positive
and negative control of the cell cycle
2. genes those involved in the repair of DNA
mismatches (initially identified in colorectal
cancer)
Cyclins and Cyclin-Dependent Kinases
Transition from one stage to the next in the cell
cycle :
regulated at a number of checkpoints at which
the integrity of the DNA are checked which
prevent entry into subsequent stages with
damaged DNA
carefully controlled by the sequential :
activation and degradation of the cyclins
activation of the cyclin-dependent kinases
(CDKs)
Cyclins and Cyclin-Dependent Kinases
Thirteen mammalian cyclins have been identified,
each one of which is required at a different stage
of the cell cycle
Six mammalian CDKs have been identified
Activation of the CDKs occurs by :
phosphorylation of a conserved threonine
residue (at position 160)
binding of the cyclin
Mammalian Cyclins and Cyclin-Dependent Kinases
Cyclin Family CDK Stage
A Mitotic CDC2, CDK2 S, G1, M
B 1-3 Mitotic CDC2 M
C G1 ? ?
D 1-4 G1 CDK2,4,5,6 G1
E G1 CDK2,4,5,6 G1/S
F ? ? ?
G ? ? ?
H ? p40 mo15 ?
The Stages of the Cell Cycle and
Expression of the Cyclins and CDKs
Cyclin A and CDC2
Cyclin B and CDC2 M
G2
G1
Cyclin D and
CDK2,4,5,6
S
Cyclin A1 Cyclin E + CDK2
CDK2
Cyclin-Dependent Kinase Inhibitors
Control of cyclins and CDKs is now also known to
occur via a group of inhibitor proteins known as
cyclin-dependent kinase inhibitor (CDKIs)
There are seven different CDKIs in mammalian
cells which belong to two different classes
Inhibitor Target Chromosomal location Regulator
p15 CDK4,6 9p21 TGF
p16 CDK4,6 9p21 ?
p18 CDK4,6 1p32 ?
p19 CDK4,6 19p13 ?
p21 CDK2,3,4,6 6p21 p53, TGF
p27 CDK2,4,6 12p12-13 Rapamycin
cAMP
p57 CDK2,3,4 11p15 ?
Control of the Cell Cycle
Each of the cyclin-CDK complexes, together with
the CDKIs :
responsible for controlling different stages of
the cell cycle
by preventing progression through checkpoints
in the presence of DNA damage
deregulation of these processes has been
implicated in tumorigenesis
1. START
The cyclin-CDK complexes linked to the regulation
of START are the D type cyclins
There are four cyclin Ds : D1, D2, D3 and D4
expressed in a cell lineage-specific manner
synthesized in response to growth factors
very short lived, rapidly degraded when growth
stimuli are withdrawn regardless of the position
of the cell cycle
if removed during G1, cells will not enter S
phase
Deregulation of cyclin D synthesis :
make cells less dependent on growth stimuli
likely to contribute to tumorigenesis
Overexpression of cyclin D1 is associated with
esophageal, breast and gastric cancers
CDK4 : has a potential role in tumorigenesis
a target for TGF in some cells
CDKIs : involved in tumor development at this
stage of the cell cycle
p16, encoded by the CDKN2 or MTS1
gene, is an inhibitor of CDK4
2. G1 to S Phase
Cells which have suffered DNA damage :
prevented from entering S phase
blocked at G1
this process is dependent on :
the tumor suppressor gene, p53
the cyclin-dependent kinase inhibitor, p21
Increased p21 Bind to a number of
cyclin-CDK complexes :
cyclin D-CDK4
cyclin E-CDK2
Activation of p53
cyclin A-CDK2
DNA damage Prevents phosphorylation of RB
Cell cycle arrest in G1
Mode of G0
RB
No transcription
E2F DP-1
Action of RB
RB
G1 E2F DP-1
No transcription
Cyclin D1/CDK4-6
Transcription
Cyclin E/CDK2 P P
RB E2F DP-1
S
TTTCGCGC
P
Cyclin A/CDK2 During G1, cyclin
P P Phosphorylation of RB
D1/CDK4-6 and cyclin
RB During
is maintained
G0 and byG1 cyclin
RB is
G2 E/CDK2 phosphorylate
underphosphorylated
A/CDK2 until mitosis
RB and E2F-1 is
P and isitbound
when is to the
released to interact
E2F-1 transcription
dephosphorylated
with and promote
factor either
ready complexed
to re-
with
Dephosphorylation M RB transcription from
DP-1. G1 or to go into
enter
genes necessary for S
the stationary phase.
phase.
RB
G0 G1 E2F DP-1
No transcription
3. S Phase
Cyclin A :
expressed from S phase through G2 and M
binds to two different CDKs :
complexed to CDK2 (during S phase)
complexed to CDC2 (during G2 and M)
has a role in both transcriptional regulation and
replication
binds to the E2F transcription factor
one of the first cyclins to be implicated in
tumor development
The cyclin A gene was the unique insertion site for
the hepatitis B virus in one clonal tumor
The virus integrate into the second intron of the
gene production of a chimeric protein (in
which the region, N terminal to the cyclin box,
was replaced with viral sequences) removal of
the ‘destruction’ box necessary for the
degradation of the cyclin in mitosis
4. Mitosis
Entry to mitosis is signaled by the activation of
the cyclin B-CDC2 complex
This complex accumulates during S and G2 but is
kept inactive by phosphorylation of tyrosine 15 and
threonine 14 residues
Mismatch Repair Genes
Mismatch Repair Genes
Both sporadic and hereditary colorectal cancers
shows defect in these genes
Hereditary nonpolyposis colon cancer (HNPCC)
accounts for around 10-15% of all colorectal
cancer
The causative gene was mapped to chromosome
2p21 by linkage studies
When DNA from tumors was compared with DNA
from normal tissues :
tumor DNA showed widespread alterations in
short repeated sequences distributed
throughout the genome
seen as additional bands over and above the
usual one or two alleles identified in the normal
tissue DNA
replication errors, caused by slippage of DNA
polymerase, had occurred during tumor
development and had not been repaired
Mismatch Repair Genes
Bacteria Yeast Human
MutS MSH2 hMSH2
GTBP
MutL MLH1 hMLH1
MutL PMS1 hPMS1
MutL PMS2 hPMS2
DNA
lesion
APOPTOSIS
Tumor
Suppressor
genes
DNA repair
STOP
G0
Apoptosis
Apoptosis (Kerr et al., 1972)
Program Cell Death (PCD)
Genetically controlled unwanted cell
- during morphogenesis
- during proliferation and differentiation
critical point of cellular control
modulated physiologically (itself & its environment)
normal regulation
self destruction mechanism
Characteristic of Apoptosis
Apoptotic cells are recognized &
phagocytosis
Differs from necrosis / accidental cell death
- active process
- no surrounding tissue damage or
induction of inflammatory responses
Apoptosis
Characteristic morphology :
- condensation of nuclear heterochromatin
- cell shrinkage
- loss of positional organization of organellas in
cytoplasm
- ladder phenomen
Controlled by protooncogenes & tumor suppressor
genes
Protein Controlling Apoptosis
Promoting Inhibiting
Intrinsic p53 BCL2/BCLXL
MYC A20
Interleukin-1
converting enzyme
BAX/BCLXS
Extrinsic TNF Many, for example :
TGF erythropoietin
PDGF/IGF1
sex hormones
Therapeutic induction of
apoptosis
• Haemopoietic malignancies
(responsive therapy) - Apoptosis >>
• high expression bcl-2 - inhibit the cell
- apoptotic responsiveness to
dexamethazone, methotrexate,
etoposide, vincristine, cisplastin &
Cyclophosphamide
• Mechanism of drug resistance ?
APOPTOSIS NECROSIS
Physiological or
Always pathological
pathological
Single cell Sheets of cells
Energy dependent Energy independent
Cell shrinkage Cell swelling
Membrane integrity Membrane integrity
maintained lost
APOPTOSIS NECROSIS
Role of mitochondria & No role for mitochondria
cytochrome c
No leak of lysosomal Leak of lysosomal
enzymes enzymes
Characteristic nuclear Nuclei lost
changes
Apoptotic bodies form Do not form
DNA cleavage No DNA cleavage
Activation of specific No activation
proteases
Regulatable Not regulated
APOPTOSIS NECROSIS
Evolutionarily Not conserved
conserved
Dead cells ingested Dead cells ingested
by neighboring by neutrophils
cells ¯ophages
Interaction of the cyclins
DNA damage
TGF beta
p53
p16 Cyclin E Cyclin A & B
p21 p27
Cyclin D proteolysis proteolysis
degradation
Cyclin D Cyclin E Cyclin A Cyclin A Cyclin B
CDK4 CDK2 CDK2 CDC2 CDC2
serum RB
E2F cdc25A
CDC25c
DNA synthesis
START S PHASE G2 PHASE MITOSIS
