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Cdc28
For S
Phosphorylate Cln1/Cln2 Clb14 expression
Regulate replication Other events for G2/M
SCF binding initiation at origins
Degradation of Cln1/Cln2
Turnoff G1 events
G1
S
Segregation of Sisterchromatids
G1 Replication Mitosis
Sister Chromatid Separation does
not Depend on the Mitotic Spindle
Cohesin:
SMC1
SMC3
SCC1
SCC3
Etc.
Cohesion is established during
replication
Dissociation of Cohesin in Prophase
Polo kinase phosphorylates cohesins and
dissociate them in chromosome arms.
Centromere (CEN1) of fission yeast
requires chromatin silencing
SWI6=heterochromatin protein 1; rad21=SCC1
Methylation and Acetylation of
Histonesthe histone code
methylation
acetylatio
n
RNAi Machinery is Required for
Chromosome Separation
RNAi machinery is required for SCC1/Cohesin
and HP1/Swi6 binding to centromeres
centromere
Histone methylation
Heterochromatin centromere
HP1/Swi6 Cohesin binding
Mitotic cyclins
(cyclin A and cyclin Byeast Clb14)
Prevent replication
origin licensing Activate APC/C Chromosome packaging
Nuclear membrane
Breakdown,
Etc.
Inhibit Sphase Chromosome
and rereplication segregation
AnaphasePromoting Complex/Cyclosome
ubiquitin E3 ligase (APC/C) complex:
a major regulator of mitosis
APC protein complex contains:
CDC27, APC11, APC4, APC13, CDC26, CDC20 (or CDH1),
AMA1, CDC23, MND2/APC15, CDC16, APC9, APC10, APC2,
APC1, and APC5.
Cyclin B/CDC2 PO4
APC APC
PO4
Inactive Active
Cleavage of SCC1 at centromere
by Separin in mitosis
centromere
Separin=Esp1
Securin=Pds1
Condesin:
SMC2
SMC4
CAPD2
CAPG
CAPH, etc.
form chromosome
loops for high order
structure in mitosis
Cyclin A Suppresses the Origindependent
Chromosome Reduplication
CDC6 CDT1 CDC6 CDT1
ORC MCMs ORC MCMs
mitosis Cyclin A
Loss of Cyclin A but not Cyclin B
Induces Polyploid Nuclei
A D
Control
B E
Cyclin A KO
C F
Cyclin B KO
1 day 2 days
KO KO
8N polyploid cells
Spindle Checkpoint in Mitosis
Taxol, Nocodazole, Colchicine
(cohesin cleavage)
Mitotic cyclins are destroyed
by APC/C ubiquitin E3 ligase
APC/C binding
CycB Cyc B
CycB CycB
The MitoticExit Network (MEN)
Nuclear Nucleolus
CDC14 in mitoticexit
Growth factor
Signaling
In mammals
Cell Cycle Regulation/Differentiation
The Mammalian Cell Cycle
Cyclin B/CDC2
Cyclin D/CDK4/6
Cyclin A/CDK2
Cyclin E/CDK2
Cell cycle in mammalian cells
Mammalian has multiple cyclindependent kinases (CDKs):
CDC2/CDK1: only for G2/M
CDK2: G1/S and G2
CDK4 and CDK6: G1
CDK3: may be similar to CDK2 but the level is very low
CDK5: central role in neuron/nerve system
Isolated by complementing yeast cdc2/cdc28 mutants, PCR,
or antibody crossreactivity of yeast CDC2.
Mammalian cyclins
Cyclin A: Sphase and G2/M
Cyclin B: mitosis
Cyclin E: G1/S transition
Cyclin D: G1
Cyclin D and cyclin E are called G1 cyclins:
complement yeast G1 cyclin (Cln13) deficient mutants
critical for mammalian G1 progression and G1/S transition
Cell cycle and cancer
Cyclin D:
15% of breast cancer has cyclin D1 gene amplification
Translocated in lymphomas and parathyroid cancers
Overexpressed in 5075% breast cancers, as well as in other
cancers
Cyclin E:
elevated level is observed in a variety of carcinomas including
breast, ovary, endometrium, stomach, colon, and acute lymphocytic
leukemias.
CELL CYCLE CONTROL BY CDKS AND CDK INHIBITORS
Cyclin B/CDC2 M
Cyclin D/CDK4 & 6
Cyclin A/CDK2 S
p16Ink4a, p15Ink4b, p18Ink4c, and p19Ink4d:
binds only to CDK4 or CDK6
inhibits cyclin D/CDK4/6 activity
The p16Ink4A gene is located on chromosome 9p21, its
loss is associated with familial melanomas and
pancreatic carcinomas
Isolated as CDK4 interaction proteins
Mammalian CDK inhibitors:
the p21/p27 family
The p21Cip1, p27Kip1, and p57Kip2:
binds to the binary complexes of G1 cyclins:
cyclin D/CDK4/6, cyclin E/CDK2
also binds to cyclin A/CDK2 and cyclin B/CDC2
isolated as CDK binding proteins
Retinoblastoma susceptibility
gene product (Rb, Rb105)
located on chromosome 13q14
Loss of heterozygosity causes retinoblastomas in
children
tumor
Rb107, Rb130, and Rb105 form the Rb family.
bind to E2F transcription factors and suppress E2F activity
P53: the most mutated tumor
suppressor in human cancers
More 50% human cancers have p53
mutation
Regulates the expression of p21 CDK
inhibitor for cell cycle arrest
Regulate proapoptosis proteins (Bax,
etc.) to induce apoptosis.
Cyclin E
Pol alpha
PCNA
Etc.
CDK inhibitors and cancer
Loss of p16Ink4a/p19ARF at 9p21 in familial melanomas and
many other cancers
Loss of p53 and Rb in human cancers
Loss of p27 is associated with malignant human cancers and serves
as best prognostic marker for breast and other cancers
p21 and p16 are associated with cellular senescence or cell aging
Regulation of replication origin firing
by cell cycle regulators
CDC6 CDT1
G1 ORC MCMs
Prereplication Origin
Cyclin E and cyclin A
S
Regulation of cyclin expression
in the cell cycle
G1 cell cycle regulation
Nutrients, Growth factors,
Cell attachment, etc.
Cyclin D Expression:
Cyclin A
G1 G1/S S G2/M
PO4
p27Kip1
p27Kip1 CDK2
Cyclin E
CDK2
inactive active
Cyclin E
How cells sense the cell mass and external
signals to degrade p27 in late G1?
Mitotic regulation in G1 (APC/CCDH1)
G0 G1 Late G1
Elevation of SKP2 is Associated
with Many Human Cancers
SKP2 as the most clinically significant
marker for human prostate cancers (not p53,
Rb, cyclin D)
elevation associated with premalignant and
malignant prostate cancers (6080%)
associated with PSA, Gleason score, and
p27 downregulation
Regulation of p27Kip1, p21Cip1, and
cyclin E by SCF ubiquitin E3 ligases
CUL1 CUL1
SKP1 SCF SKP2
SKP1 SCFFBW7/hCDC4
FB SKP2
FB FBW7
PO4 PO4
p27 Cyclin E
PO4 Notch, Myc, etc.
p21
FB: Fbox
More than 40 Fbox proteins exist
p16Ink4a and p19ARF are both tumor suppressors and
are located on overlapping sequences on 9p21
ARF:
Alternative Reading Frame
Cell Cycle Regulation
APC/CCDC20/CDH1
Cyclin B/CDC2
DNA damage
DNA damage
Replication MDM2
Cyclin A/CDK2 Cyclin D/CDK4/6
p27 and p21 p53
Cyclin E/CDK2
SCFSKP2 E3 Ligase
Growth factors and cell
attachment
DNA damage response in mammals
p21
Why mitosis does not occur in S phase cells?
ATR/CHK1
CDC25C
G2/M
DNA damage response in the cell cycle
PO4
C Cdc25A
SCFβTRCP
Cdc25A
G2 arrest
CDK2
G1 arrest
G1 arrest
The G2 checkpoint
Genome stability and cancer
polyploidy and aneuploidy
chromosomal deletion, gene amplification, translocation
increased mutational rate
Defects in the cell cycle checkpoints
Mitosis versus Meiosis
(SCC1)
+ Polo
+ Polo
(Rec8)
asymmetric cell division and cell fate
Asymmetric cell division
and cell fate determination
CCCH Proteins
CCCH Proteins
Differential
proteolysis of
CCCH finger
proteins by cullin 2
in germ and
somatic cells
To keep the germ
cell fate in C. elegans
Future direction:
How to cure human diseases
such as cancer and proliferation
or agerelated diseases?
Mechanisms
Disease genes
Therapeutic strategies
Regulation of Rb proteins in G1