Studies of the interaction between Rad52 protein and the yeast single-stranded DNA binding protein RPA
Studies of the interaction between Rad52 protein and the yeast single-stranded DNA binding protein RPA
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DOI：
10.1128/mcb.18.7.4400
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发表时间：
1998-07-01
影响因子：
5.3
通讯作者：
Berg, P
中科院分区：
文献类型：
--
作者：
Hays, SL;Firmenich, AA;Berg, P
RFA 1基因编码酵母三聚体单链DNA结合蛋白复制蛋白A（RPA）的大亚基，已知其在DNA复制中起关键作用。携带rfa 1 -44等位基因的酿酒酵母菌株显示出许多受损的重组和修复表型，所有这些表型都可以通过过表达RAD 52来抑制。我们证明，rad 52突变是上位的rfa 1 -44突变，将RFA 1和RAD 52在相同的遗传途径。此外，双杂交分析表明Rad 52和RPA的所有三个亚基之间存在相互作用。通过使用rad 52的两种不同的突变等位基因进一步探索了这种Rad 52-RPA相互作用的性质。两种突变都位于Rad 52的氨基末端，这是先前定义为负责其DNA结合能力的区域（U，H，Mortenson，C，Beudixen，I，Sunjeuaric，和R，罗斯坦，Proc.Natl.科学院USA 93：10729-10734，1996）。酵母双杂交系统用于监测突变体Rad 52蛋白的蛋白-蛋白相互作用。这两种突变蛋白都具有自身相互作用的能力，但不能与Rad 51相互作用。突变蛋白也缺乏与RPA大亚基Rfa 1相互作用的能力。有趣的是，它们保留了与中等大小的亚基Rfa 2相互作用的能力。鉴于Rad 52的DNA结合结构域中的突变的位置，提出了一种模型，该模型将DNA在DNA双链断裂修复中所涉及的蛋白质-蛋白质相互作用中的作用结合起来。
The RFA1 gene encodes the large subunit of the yeast trimeric single-stranded DNA binding protein replication protein A (RPA), which is known to play a critical role in DNA replication. A Saccharomyces cerevisiae strain carrying the rfa1-44 allele displays a number of impaired recombination and repair phenotypes, all of which are suppressible by overexpression of RAD52. We demonstrate that a rad52 mutation is epistatic to the rfa1-44 mutation, placing RFA1 and RAD52 in the same genetic pathway. Furthermore, two-hybrid analysis indicates the existence of interactions between Rad52 and all three subunits of RPA. The nature of this Rad52-RPA interaction was further explored by using two different mutant alleles of rad52, Both mutations lie in the amino terminus of Rad52, a region previously defined as being responsible for its DNA binding ability (U, H, Mortenson, C, Beudixen, I, Sunjeuaric, and R, Rothstein, Proc. Natl. Acad, Sci. USA 93:10729-10734, 1996), The yeast two-hybrid system was used to monitor the protein-protein interactions of the mutant Rad52 proteins. Both of the mutant proteins are capable of self-interaction but are unable td interact with Rad51, The mutant proteins also lack the ability to interact with the large subunit of RPA, Rfa1. Interestingly, they retain their ability to interact with the medium-sized subunit, Rfa2. Given the location of the mutations in the DNA binding domain of Rad52, a model incorporating the role of DNA in the protein-protein interactions involved in the repair of DNA double-strand breaks is presented.