ADP stabilizes the human Rad51-single stranded DNA complex and promotes its DNA annealing activity

被引:24
作者
Kim, HK
Morimatsu, K
Nordén, B
Ardhammar, M
Takahashi, M
机构
[1] Chalmers Univ Technol, Dept Chem Phys, SE-41296 Gothenburg, Sweden
[2] Univ Paris 11, Inst Curie, F-91405 Orsay, France
[3] Univ Paris 11, CNRS, F-91405 Orsay, France
[4] Univ Nantes, F-44322 Nantes 3, France
[5] CNRS, FRE 2230, F-44322 Nantes 3, France
关键词
D O I
10.1046/j.1365-2443.2002.00588.x
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Background: Human Rad51 protein (HsRad51) is a homologue of Escherichia coli RecA protein, and involved in homologous recombination. These eukaryotic and bacterial proteins catalyse strand exchange between two homologous DNA molecules, each forming a complex with single-stranded DNA (ssDNA) and ATP as the initial step. Both proteins hydrolyse ATP; however, the role of ATP hydrolysis appears to vary between the two proteins. Results: Measurements using the fluorescence ssDNA analogue, poly(1,N (6) -etheno-deoxyadenosine), indicate that ATP affects the HsRad51-ssDNA complex, promoting two conformational states: one transient, rather rigid transition state and a final more flexible state. While ADP lowers the affinity of RecA protein to ssDNA, it is found to rather stabilize the HsRad51-ssDNA complex. ADP does not activate the strand exchange by HsRad51 but instead stimulates annealing between complementary ssDNAs. Conclusions: The hydrolysis of ATP promotes a transition of the HsRad51-ssDNA complex from a stiff state to less stiff state. The first state may be important for the strand separation of dsDNA in the initial step of strand exchange, while the second state may be important for annealing in the next step. However, hydrolysis does not dissociate HsRad51 from DNA as a component step of its recycling.
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收藏
页码:1125 / 1134
页数:10
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