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DNA repair
DNA repair is a cellular mechanism to correct damage to DNA before it becomes fixed as a mutation or chromosomal aberration, which may lead to deleterious results such as cell death or tumorigenesis. Mechanism of DNA repair is important for reducing the risk of cancer as well as developing more effective cancer therapies.
MUTASI
Mutasi terbagi atas: Mutasi spontan Mutasi induksi
Direct repair
Observation: UVPhotoreactivation
requires DNA photolyases requires visible light at 300 500 nm light repair Contrast: dark repair (BER, NER, mismatch repair)
DNA photolyases
Structure
Generally contain 2 chromophores Chromophore No. 1: always FADH Chromophore No. 2: folate (in E. coli and yeast)
N5,N10-methenyltetrahydrofolylpolyglutamate
Function
bind to pyrimidine dimers resolve pyrimidine dimers into original bases
UV-responsive photolyases
DNA glycosylases
Relatively small P enzymes (20 30 KDa) Recognize abnormal bases
deaminated bases alkylated bases
P
G P O U
O
G
O
Remove base via cleavage at the glycosidic bond between the deoxyribose and the base Cleavage creates apurinic and apyrimidinic (AP sites)
A
O
A
O
Before
After
AP-endonucleases
5 P P P P P P P recognize AP-sites A G G C A G C cleave phosphodiester bonds near the AP site T C C T C G andgeneratea5 3 P P P P P P P phosphateand3AP endonuclease hydroxyl In E. coli this enzyme alsohas5-3 P P P P P P P exonuclease activity A G G C A G C The-3OH functions as T C C G a primer 5 P P
3
4. ligation of the break in the strand. Two enzymes are known that can do this; both require ATP to provide the needed energy.
Function
scans DNA, binds to UvrB scanner; binds DNA cleaves phosphate bond at 3' end, 5 positions downstream of lesion binds UvrB & DNA cleaves phosphate bond at 5' end, 8 positions upstream of lesion removes DNA fragment fills emerging gap seal nick
UvrB
exinuclease P
ATP
P
P
OH DNA pol. I
DNA ligase
MISMATCH
Mismatch repair
ATP ADP+Pi
5 3
CH3
3 5
CH3
3 5
ATP ADP+Pi
5 3
CH3
3 5
5 3
CH3
3 5
SOS Response
SOS repair occurs when cells are overwhelmed by UV damage - this allows the cell to survive but at the cost of mutagenesis. SOS response only triggered when other repair systems are overwhelmed by amount of damage so that unrepaired DNA accumulates in the cell.
SOS-response
Inaccurate repair mechanism Requires at least 14 proteins in E. coli Din proteins (damage induced) Rec poteins (recombination) Umu proteins (UV-mutagenesis) Uvr proteins (UV-resistance) Others: SulA, HimA, Ssb, and PolB
THANKS.