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首页> 外文期刊>Molecular and Cellular Biology >Identification of a protein that binds to the Ho endonuclease recognition sequence at the yeast mating type locus.
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Identification of a protein that binds to the Ho endonuclease recognition sequence at the yeast mating type locus.

机译：在酵母交配型基因座上鉴定与Ho核酸内切酶识别序列结合的蛋白质。

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摘要

Mating type switching in Saccharomyces cerevisiae initiates when Ho endonuclease makes a site-specific double-stranded break at MAT, the yeast mating type locus. To identify other proteins involved in this process, we examined whether extracts prepared from ho- mutants contain additional factors that bind near the recognition sequence for Ho. Using an electrophoretic mobility shift assay, we isolated a chromatographic fraction that contains an activity, named YZbp, which binds to two sequences flanking the recognition sequence at MATalpha and to one sequence overlapping it at MATa. MAT plasmids carrying mutations in the YZbp recognition sequence are cleaved by purified Ho at wild-type efficiencies in an in vitro assay. These same plasmids, however, are not cleaved by Ho inside cells, demonstrating that YZbp acts as a positive activator of in vivo cleavage. YZbp is present in all cell types, even those not undergoing mating type switching, suggesting that it has additional cellular functions.
机译：当Ho内切核酸酶在酵母交配型基因位点MAT上发生位点特异性双链断裂时，酿酒酵母中的交配类型转换开始。为了鉴定参与此过程的其他蛋白质，我们检查了从ho突变体制备的提取物是否包含在Ho识别序列附近结合的其他因子。使用电泳迁移率变动分析，我们分离了一种色谱级分，其中包含一种名为YZbp的活性，该活性与MATalpha识别序列侧翼的两个序列以及MATa重叠的序列相结合。在体外测定中，以野生型效率通过纯化的Ho切割在YZbp识别序列中携带突变的MAT质粒。然而，这些相同的质粒并未被细胞内的Ho切割，表明YZbp充当体内切割的阳性激活剂。 YZbp存在于所有细胞类型中，甚至包括那些未进行配对类型转换的细胞，这表明它还具有其他细胞功能。

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