Cell cycle regulation of DNMT1 and cancer

1
Cell cycle regulation of DNMT1 and cancer
2
The Genome and Epigenome Must be Faithfully
Replicated with Each Cell Cycle
M
M
M
M
DNMT1
M
M
M
M
M
M
M
M
M
M
G0/G1
S
G2
M
DNMT1 transcription
Enzymatic activity
Steady state mRNA
3
Expression of DNMT1 mRNA is regulated with the
cell cycle
4
The DNMT1 3UTR Contains A Highly Conserved
Element
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daniorarioutr(2342-2364)
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c
hic
k
enutr(4936-4991)
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-
-
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G
T
G
T
A
G
T
A
C
T
T
T
G
T
G
C
A
T
T
C
T
G
mouseutr(5076-5106)(5194-5233)
T
G
A
T
T
T
A
G
T
G
A
T
C
A
A
A
T
T
G
T
G
C
A
G
T
A
C
T
T
T
G
T
G
C
A
T
T
C
T
G
humanutr(5089-5408)
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C
T
T
T
G
T
G
C
A
T
T
C
T
G
rattusutr(5072-5093)(5123-5211)
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daniorarioutr(2342-2364)
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c
hic
k
enutr(4936-4991)
A
A
T
T
T
T
A
A
-
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mouseutr(5076-5106)(5194-5233)
G
A
T
T
T
T
A
A
A
A
G
T
T
T
T
T
T
A
T
T
A
T
G
C
A
T
T
A
T
A
T
C
A
A
A
T
C
T
A
C
humanutr(5089-5408)
A
A
T
T
T
T
A
A
-
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-
T
T
T
T
T
T
A
T
T
C
T
G
T
A
T
T
C
T
A
T
C
A
G
A
T
C
T
G
C
rattusutr(5072-5093)(5123-5211)
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G
T
T
T
T
T
A
T
daniorarioutr(2342-2364)
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A
C
T
T
T
A
T
G
T
A
G
T
T
T
T
T
A
T
c
hic
k
enutr(4936-4991)
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A
G
A
C
T
T
G
A
T
G
T
A
G
T
T
N
T
T
A
T
mouseutr(5076-5106)(5194-5233)
C
A
C
T
G
T
A
T
G
A
G
T
G
G
A
A
A
T
T
A
A
G
A
C
T
T
T
A
T
A
rattusutr(5072-5093)(5123-5211)
daniorarioutr(2342-2364)
c
hic
k
enutr(4936-4991)
mouseutr(5076-5106)(5194-5233)
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
humanutr(5089-5408)
rattusutr(5072-5093)(5123-5211)
5
A Specific Protein (p40) Binds the Conserved
Region of the DNMT1 3UTR in Arrested Cells
3UTR probes /
competitors
3
5
UTR
5090-5408
5- 259
UTR
D
5349-5405
3- 56
5090-5350
UTR
D
5090-5248
3- 158
UTR
D
serum induction
(h)
UTRD 5 259
UTRD 3 158
UTRD 3 56
36
competitor
16
20
24
0
8
none
UTR
kDa
130
85
43
32
6
a
a
a
a
a
a
a
DNMT1 3UTR destabilizes mRNA in arrested cells
serum arrested
?0 .5 1 2 4?
?0 .5 1 2 4?
56 nt
1
t1/2105 min
0.1
t1/258 min
0.01
??
0
100
200
300
7
The Highly Conserved Region of the 3UTR is
Required to Downregulate the DNMT1 mRNA in
Growth Arrested Cells
serum induced (20h)
growth arrested
D
D
DNMT1
UTR
D

5085
0
DNMT1
DNMT1-UTR
56
D
5350
DNMT1
5408
GFP
CMV
GFP
30
polyA
pAD-DNMT-1
25
20
growth arrested
Normalized DNMT1 mRNA
Kan
15
serum induced (20h)
10
5
0
DNMT1
DNMT1
DNMT1
UTR
D
8
Deregulated cell cycle expression of DNMT1
leads to cellular transformation
NIH 3T3
h-DNMT1
h-DNMT1-D3UTR
9
The 3UTR inhibits ectopic expression of DNMT1 at
the G0 phase of the cell cycle
Ectopic expression of DNMT1 induces entry into
the S phase of the cell cycle
10
Mitogenic signals
DNA replication
DNA replication
DNA methylation
Protection of the DNA methylation pattern
Fig. 2
11
Inhibition of DNMT1 by antisense oligonucleotides
inhibits Y1 tumor growth in vivo
12
(No Transcript)
13
Nancy Detich QiangLi Zhuang Nadia Cervoni Ian
Weaver Michael Meany