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首页> 外文期刊>Molecular and Cellular Biology >Stable and specific association between the yeast recombination and DNA repair proteins RAD1 and RAD10 in vitro.
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Stable and specific association between the yeast recombination and DNA repair proteins RAD1 and RAD10 in vitro.

机译：体外酵母重组与DNA修复蛋白RAD1和RAD10之间的稳定和特异性结合。

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摘要

The RAD1 and RAD10 genes of Saccharomyces cerevisiae are two of at least seven genes which are known to be required for damage-specific recognition and/or damage-specific incision of DNA during nucleotide excision repair. RAD1 and RAD10 are also involved in a specialized mitotic recombination pathway. We have previously reported the purification of the RAD10 protein to homogeneity (L. Bardwell, H. Burtscher, W. A. Weiss, C. M. Nicolet, and E. C. Friedberg, Biochemistry 29:3119-3126, 1990). In the present studies we show that the RAD1 protein, produced by in vitro transcription and translation of the cloned gene, specifically coimmunoprecipitates with the RAD10 protein translated in vitro or purified from yeast. Conversely, in vitro-translated RAD10 protein specifically coimmunoprecipitates with the RAD1 protein. The sites of this stable and specific interaction have been mapped to the C-terminal regions of both polypeptides. This portion of RAD10 protein is evolutionarily conserved. These results are the first biochemical evidence of a specific association between any eukaryotic proteins genetically identified as belonging to a recombination or DNA repair pathway and suggest that the RAD1 and RAD10 proteins act at the same or consecutive biochemical steps in both nucleotide excision repair and mitotic recombination.
机译：酿酒酵母的RAD1和RAD10基因是至少七个基因中的两个，已知它们是核苷酸切除修复期间DNA的损伤特异性识别和/或损伤特异性切口所需要的。 RAD1和RAD10也参与专门的有丝分裂重组途径。我们先前已经报道了将RAD10蛋白纯化至同质性（L.Bardwell，H。Burtscher，W.A.Weiss，C.M.Nicolet，和E.C.Friedberg，Biochemistry 29：3119-3126，1990）。在本研究中，我们显示了通过克隆基因的体外转录和翻译产生的RAD1蛋白，与体外翻译或从酵母中纯化的RAD10蛋白共免疫沉淀。相反，体外翻译的RAD10蛋白与RAD1蛋白共免疫沉淀。这种稳定和特异性相互作用的位点已被定位到两个多肽的C-末端区域。 RAD10蛋白的这一部分在进化上是保守的。这些结果是遗传鉴定为重组或DNA修复途径的任何真核蛋白之间特异性结合的第一个生化证据，表明RAD1和RAD10蛋白在核苷酸切除修复和有丝分裂重组中以相同或连续的生化步骤起作用。

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