'

7
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- lga1@bezeqint.net

30.11.2004
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The Cell-Cycle in Eukaryotic Cells

G1
. S-' G1- DNA .
S- DNA- gap G2 M- .
G1 '
. .

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G1-
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-DNA- -
.

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DNA .

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. Rb
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. S-
, " ' DP' . E2F
G1- E2F-DP .S-
Rb-" E2F-DP
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Rb- AD- BD- Rb , AD-
DNA- - DNA-.
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Rb ' . G1
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Rb Rb -
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7.12.04

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10% .brca1
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.
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CDK .RB ,
.. g1 .
RB .
.rb g1 cdk4 "
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cdk2 E . 2'
.g1

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RB . CDK2 .
. RB
.RB
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. p16 .KD 16 cdk4,6
p16 , .
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P16 , 2- , 50%- .
P21 . ,DNA .
p21 .p53 " ".
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DNA Replication Depends on the Activation of an Origin of Replication

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14.12.2004

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S-
origin of replication
.
origin of replication
.
origin of replication
.

11

Origin of Replication- General Structure

origin of replication- 200-

.

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, origin of replication - .core- origin ORC
.Recognition complex
- ORC -Origin regulatory elements .ORC origin of
.replication
ORE - 2 , . .T,A
.
.
DNA .
. DNA
.
dna unbiding element -DUE .
ORC .
origin of replication . core-
. origin of replication- .
. transcription factors
.RNA DNA .origin of replication -
.DNA-

 '

12

Transcription and DNA Replication

,tata box origin of replication- origin .ORE

Recognition complex .
.DNA
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,ORC
" origin of replication - .
.DNA origin of replication
. S- " .
S- .
" .
origin of replication S- .S-
S- "
.
.origin of replication
origin of replication ,
. origin of replication
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S- . . S-
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13

origin of -
replication .
,origin of -
, replication ,
. ? origin of replication-

. origin of replication-
. sv40 .T
RB .S "
".
T - , ,
,RB- origin of replication-
.
ORC intiation proteins ,
origin of replication - . T
.
: ,
.S
.

 '

14

licensing

origin of replication-?
" " licensing
?
.

G0 origin of replication
. ORC . origin of replication ORC origin of -
.replication . G0-
ORC 2 , Cdc6 .
.cell devision control = Cdc .
, cdt1 . " ORC- origin
.of replication
origin of replication - . S-
, MCM .mini chromosome maintaining proteins
MCM cdt1- cdc6- .DNA- MCM
, . MCM
.DNA-
S- MCM- . S- origin of replication
.MCM-
origin of replication . origin of replication -
.ORC- cdt1- cdc6- .
. MCM
S- cdt1 cdc6- .ORC - ? 2.
. , Geminin ,S
cdt1 .ORC
Cdc6 S , cdc6 .
' . cdt1 cdc6-
.

 '

15

3:
cdt1 cdc6- S-
cdt1 cdc6-
cdt1cdc6-
' cdt1 cdc6- .
" .
, cdt1 cdc6- ORF
. ' ,cdt1 -
' .
cdt1 cdc6- . origin of replication-
' cdt1 . cdc6-
.DNA- ' - MCM
.
' : origin of replication
cdt1 cdc6- .
-' .cdc6

 '

16

21.12.2004
" " " .
10 DNA . leading 5- 3-
, lagging -
RNAase " .
200 .
/ 2 " .

Replication of the Leading and Lagging Strands

.RNA DNA DNA . .0-

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5- 3- . lagging strand

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5- 3- ? ?
. .
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.

17

Alternative Replication Model

3- .5- 5 3- .
, . .
. 5- 3- ?
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3- ,5- .
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5- 3- . lagging strand
" leading strand .
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.

Processivity of DNA Polymerases

 '
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leading lagging- .
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ATP .
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.

lagging
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.SSBP
leading
. lagging
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2' .
. lagging
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" lagging
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DNA Replication Folk.

The Replication Complex Model

18

 '
" leading , lagging .
.

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19

" ATP
.
" , SV40 6000.
" , leading
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PCNA " . .
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RNA ,
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DNA
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, leadung
lagging .

 '

20

MISMATCH
DNA .
2 - .
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.

' mismatch reapair " . 2

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. ,
.

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21

' .
. MuTH, MutL ,MutS " .
' ' 2 .
MutL , , nick mismatch.
.mismatch 1
3- ,5- 7 5-.3-

1 7 , 3 .
.

 '

22

28.12.2004
.
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2- .
RNA . .
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2 2 . 3 .
?3 ' 5-.3-
DNA- . . lagging
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S- , S- .
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Extension of Telomers in Eukaryotic Cells

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S- 2
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Trf1 , Trf2
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Trf1 , Trf2- .
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" .Trf1 , Trf2 ? .

23

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2 , 2
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P53
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P53 .
p53 ? p53 .1979-
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 '
p53 . 50%-
.
-p53
p53
, MDM2
.
. Mdm2
Mdm2 .
p53
.


. Mdm2-

.

.

24

p53 .tumor suppressor gene p53

/ 2- . p53-
" .missense , p53
. .
p53 393' p53 . : p53 .
. dna binding domain- .transcription activation domain 2-
' 2 . 20 - 15 N-
. . 20
p53- .

 '
15 20 .
,
.p53 .
p53 ? T p53
. - .
DNA- .
T . .

25

DNA- p53- . Atm ,

.DNA- p53 15 .
Atm check2 p53 .20
, C , ,p53
.DNA
p53 4- . .DNA-

 '

26

:
: DNA-
' ATM .
p53 .DNA-
, DNA bundung donmain- p53
.
p53 DNA- " . p53-
, MDM2 , p53- , '
, p53 .
MDM2 p53 . MDM2
.
,DNA- ATM , p53 ,15
chek2 p53 ,20 MDM2 p53-
, p53 .
DNA damage
ATM/ DNA pk
MDM2

p53
Bax/Bcl2

P21 Waf1/ Cip1

Apoptosis

Cyclin D/ cdk

R
pRB + E2F

G1

Rb
E2F

G2
M

Cell cycle damage and control

p53 ? . . P21
Cdk . cycklin D .
p53 .Bax
p53 , .DNA-
p53 . .
,
.
, p53 .

 '

27

p53 " Mdm2 . Mdm2 p53- N . p53-

.
Mdm2 ,p53 .

 '
p53- ,
enhansor mdm2 . mdm2 .
p53 mdm2 " . p53 DNA-
mdm2 .20 ser-
DNA- ATM , p53 , 20
mdm2- . p53 DNA- . p53
.

28

- 3'

 '

29

4.12.2005
PRB tumor supressor .
- tumor .
supressor . .
P53 ,tumor suppressor .care taker .DNA-
care takers .
P53 DNA- , ) NLS
. .MDM
ATM- p53 p53 , .
.
- .DNA-
1G S- .
P21 4/6 CDK- - 1D 2 CDK- .E
RB G1-.S-
P53 ,
. S - p21 '
. p53 S- , GAD-45
PCNA . PCNA DNA- . PCNA -
DNA .
P53 . S- .DNA-
P53 G2.M-
DNA- , DNA-
DNA- p53.
DNA- 2 , ,
DNA .
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2 - '
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,DNA- " " .
DSB
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DN- , . -
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.DNA-
.
NIC/ .DNA- .
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.

 '
.DNA- 2- ,
.
.
.DNA- double stranded breaks -DSB .
2- , . DSB
.

30

?
S- .
-DSB .S- "
.DNA- S- .DNA DNA-"
. DSB " ' Non homolgos end .NHEJ
- joininig .
.
,NHEJ - , G1- .
, ? ,S- .
' ,DSB- ' .
, .DSB ' -
DSB . , '
.
DSB- ' .
BRCA1
care takers - .BRCA1
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BRCA1- 80% . tumor suppressors-care -
takers 2 , . ,
.
, 80% 50% ?
? BRCA1 1863" . ' . RNA 2
.
" ,ATM 2- .
, BRCA2 , 1- 3418" ,
.80%-

 '

31

? -
BRCA2 . .
RAD51 ' " .
BRCA1 .BRCA2 BRCA1 .
BRCA2 .
DSB - ,
.
DNA" ,
. .
- ' .
. 2 .
.
DSB ,DNA- ' .DSB-
ATM ATR- . CHK2- " DNA-
.
.DNA-
" . BRCA1 " " , " ,ATM ,
. . Nbs1, MRE11, RAD50

MRE11 . ' 2 ,DNA-

MRE11 2 ' - cohesive ends DSB - "".
, BRCA2 , RAD51
, REC A -.
RAD51 , -
.

 '
homolegos recpmbination .DNA polimeraz
.
, 2 .
,
, , RAD51- , " . BRCA2

32

:
BRCA1 MRE11 BRCA2 . RAD51
. BRCA1 - BRCA2- 2 .
BRCA2 , RAD51
.

BRCA1- , BRCA2 ? ? DSB

, . DNA-
. RAD51- ,
.

 '
2 BRCA- , 1BRCA ? ? DSB
.

33

BRCA1 - NHEJ- .
, BRCA1 , DSB ,
, BRCA1- .care taker
.
BRCA2- .
. 80%- .
?
BRCA1 BRCA2 - , .
.
BRCA1- ' . .
. histon de-acetylase
histon de-acetylase , 13 ' .2
histon de-acetylase .
.
. .DNA- ,
.DNA-
Brca1 , .
DSB .GAD45-
p53 , GAD45 .histon de-acetylase PCNA
. DNA DNA-.
Brca1 .
Brca1 , ,histon de-acetylase
DNA -.
.
.
S- , -
.DNA
':
.
.
) pH( . - ) pH(.
?
, DNA-
.
RNA ? U-A , T-A
RNA .stem&loop
5 stem&loop -
.RNA scaning . cap - .

 '

34

11.4.2005
DNA- , S- DNA-
.
S- DNA- ,
, ' . .
- .
. .
DNA- DNA-
.
DNA- ?
DNA- ' .
2 .
DNA- RNA . ,
DNA- .
DNA- .
- DNA- " :
.
.1 - . : -
.DNA- .
.2 - ,DNA-
.DNA-

 '

35

"
.1 UV - .
.2 - .

.
?
,
DNA- . ,
DNA- . ? ?
,
' .
?
(1 .
(2 .
' ?
BBR-
, .BBR
.
'
.
DNA-
.
.
DNA. , , .

 '
' - DNA .
.

36

. 2 .
AP .1 Apurinic endonucleaz . . 1
. NIC DNA- 2
.
.2 , ,
.
.3 3 . ?
DNA 1) DNA , ( ,
" .

?
? ? ?
AP . . 2
- . " ,
2 . .

 '
.

37

?
NER-
' . nucleotide excision repair - NER
?
.
,
, '
.DNA-
" .BBR-
.1 ) (... DNA -
.DNA- 2- ,
. .
.2 , .
DNA .3
.4 .
" 4 2 - BBR- .
.NER-
- -
.DNA DNA- .UV

 '
.E.COLI-
2 :
DNA- 2- 30 . .
ecoli - 12.
DNA ,1 2 DNA - .
?
UV , NER -
.
, .p53
p53 ,DNA- " MDM. -
. .
G1 ,S- S.M-

38