Purification and characterization of an activity from Saccharomyces cerevisiae that catalyzes homologous pairing and strand exchange.
Purification and characterization of an activity from Saccharomyces cerevisiae that catalyzes homologous pairing and strand exchange.
复制标题
酿酒酵母催化同源配对和链交换活性的纯化和表征。
DOI：
10.1073/pnas.84.16.5560
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发表时间：
1987
影响因子：
11.1
通讯作者：
Morrison,PT
中科院分区：
文献类型：
--
作者：
Kolodner,R;Evans,DH;Morrison,PT
从有丝分裂的酿酒酵母细胞中纯化1000- 2000倍的催化线性M13 mp 19复制型DNA和环状M13 mp 19病毒DNA形成接合分子的活性。该活性似乎存在于Mr 132，000多肽中。该反应要求底物是同源的，并且还需要Mg 2+。不需要ATP。该反应需要化学计量的蛋白质，并表现出合作依赖于蛋白质浓度。电子显微镜分析的关节分子表明，它们形成的单链环状分子的线性双链体的一条链的位移。该分析还显示，异源双链体形成始于线性双链体链的3 '同源末端，随后杂交区向线性双链体链的5'同源末端延伸（3 '至5'方向）。
An activity that catalyzes the formation of joint molecules from linear M13mp19 replicative form DNA and circular M13mp19 viral DNA was purified 1000- to 2000-fold from mitotic Saccharomyces cerevisiae cells. The activity appeared to reside in a Mr 132,000 polypeptide. The reaction required that the substrates be homologous and also required Mg2+. There was no requirement for ATP. The reaction required stoichiometric amounts of protein and showed a cooperative dependence on protein concentration. Electron microscopic analysis of the joint molecules indicated they were formed by displacement of one strand of the linear duplex by the single-stranded circular molecule. This analysis also showed that heteroduplex formation started at the 3'-homologous end of the linear duplex strand followed by extension of the hybrid region toward the 5'-homologous end of the linear duplex strand (3'-to-5' direction).