單分子技術觀察DNA錯誤配對辨認蛋白 MutS 如何抑制 RecA 重組酶進行部分同源股交換反應

劉泓其

doi:10.6342/NTU.2015.00043

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單分子技術觀察DNA錯誤配對辨認蛋白 MutS 如何抑制 RecA 重組酶進行部分同源股交換反應

Investigate How Mismatch Repair Protein MutS Inhibits RecA-Catalyzed Homeologous Strand-Exchange Reaction Using a Single-Molecule Method

劉泓其 , Masters　　Advisor：李弘文　　

繁體中文 DOI： 10.6342/NTU.2015.00043

RecA 重組酶；單分子栓球實驗；單分子； DNA 修復； MutS蛋白；部分同源重組反應； RecA ； TPM ； Single molecule ； DNA repair ； MutS ； Homeologous recombination

Abstract │ Reference (29) │ Altmetrics

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Homologous recombination (HR) plays an important role in meiosis and DNA damage repair. In prokaryotes, RecA nucleoprotein filaments search complementary DNA sequence, and carry out the strand-exchange reaction of essential steps in HR. Moreover, RecA can catalyze strand-exchange reaction even if DNA sequence not totally homologous, i.e. homeologous. Previous studies indicate that mismatch repair protein, MutS, inhibits RecA-mediated homeologous recombination. However, the inhibition mechanism remains unknown. We found the DNA mismatch density plays an important role in inhibiting the homeologous strand-exchange reaction. Using gel-shift electrophoresis assay, an intermediate state of strand exchange reaction with higher molecular weight appeared in the presence of MutS. This suggests that MutS proteins promote the accumulation of the reaction intermediate, thus reduce final reaction products. Using single-molecule tethered particle motion experiments, we measured interaction between RecA filaments and homeologous DNA substrates in the presence of MutS. These experiments offer information about how MutS inhibition might take place.

Reference ( 29 ) 〈TOP〉

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2 Modrich, P. & Lahue, R. Mismatch repair in replication fidelity, genetic recombination, and cancer biology. Annu. Rev. Biochem. 65, 101-133 (1996).
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4 Jeong, C. et al. MutS switches between two fundamentally distinct clamps during mismatch repair. Nature Struct. Biol. 18, 379-385 (2011).
連結：
5 Lamers, M. H. et al. The crystal structure of DNA mismatch repair protein MutS binding to a G.T mismatch. Nature 407, 711-717 (2000).
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