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酵母重组因子Rad59中的单链DNA结合与退火活性。

Single strand DNA binding and annealing activities in the yeast recombination factor Rad59.

作者信息

Petukhova G, Stratton S A, Sung P

机构信息

Department of Molecular Medicine/Institute of Biotechnology, University of Texas Health Science Center, San Antonio, Texas 78245-3207, USA.

出版信息

J Biol Chem. 1999 Nov 26;274(48):33839-42. doi: 10.1074/jbc.274.48.33839.

Abstract

Saccharomyces cerevisiae RAD59 gene is required for homologous recombination processes and normal level of resistance to ionizing radiation. To study the biochemical functions of Rad59, it was overproduced in yeast and purified to near homogeneity. Rad59 binds DNA, showing much higher affinity for ssDNA than dsDNA. Rad59 also anneals complementary DNA strands, and order of addition experiments indicate that maximal annealing efficiency is achieved when both complementary DNA strands are present upon addition of Rad59. Thus, Rad59 resembles its homolog Rad52 in being able to bind ssDNA and anneal complementary DNA strands. However, unlike Rad52, DNA annealing by Rad59 is not accelerated by the ssDNA binding factor RPA. DNA binding and strand annealing are likely to be important for the biological functions of Rad59 in general recombination and in the single-strand annealing pathway of recombination.

摘要

酿酒酵母RAD59基因是同源重组过程和正常水平的电离辐射抗性所必需的。为了研究Rad59的生化功能,它在酵母中过量表达并纯化至近乎均一。Rad59结合DNA,对单链DNA的亲和力比对双链DNA高得多。Rad59还能使互补DNA链退火,添加顺序实验表明,当添加Rad59时两条互补DNA链都存在时,可实现最大退火效率。因此,Rad59在能够结合单链DNA和使互补DNA链退火方面类似于其同源物Rad52。然而,与Rad52不同,单链DNA结合因子RPA不会加速Rad59介导的DNA退火。一般来说,DNA结合和链退火可能对Rad59在一般重组和重组的单链退火途径中的生物学功能很重要。

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