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首页> 外文期刊>Molecular and Cellular Biology >The REC2 gene encodes the homologous pairing protein of Ustilago maydis.
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The REC2 gene encodes the homologous pairing protein of Ustilago maydis.

机译：REC2基因编码玉米Us的同源配对蛋白。

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摘要

Amino acid sequence analysis has established that the homologous pairing protein of Ustilago maydis, known previously in the literature as rec1, is encoded by REC2, a gene essential for recombinational repair and meiosis with regional homology to Escherichia coli RecA. The 70-kDa rec1 protein is most likely a proteolytic degradation product of REC2, which has a predicted mass of 84 kDa but which runs anomalously during sodium dodecyl sulfate-gel electrophoresis with an apparent mass of 110 kDa. To facilitate purification of the protein product, the REC2 gene was overexpressed from a vector that fused a hexahistidine leader sequence onto the amino terminus, enabling isolation of the REC2 protein on an immobilized metal affinity column. The purified protein exhibits ATP-dependent DNA renaturation and DNA-dependent ATPase activities, which were reactions characteristic of the protein as purified from cell extracts of U. maydis. Homologous pairing activity was established in an assay that measures recognition via non-Watson-Crick bonds between identical DNA strands. A size threshold of about 50 bp was found to govern pairing between linear duplex molecules and homologous single-stranded circles. Joint molecule formation with duplex DNA well under the size threshold was efficiently catalyzed when one strand of the duplex was composed of RNA. Linear duplex molecules with hairpin caps also formed joint molecules when as few as three RNA residues were present.
机译：氨基酸序列分析已确定，Ustilago maydis的同源配对蛋白（以前在文献中称为rec1）由REC2编码，REC2是重组修复和减数分裂所必需的基因，与大肠杆菌RecA具有区域同源性。 70 kDa rec1蛋白很可能是REC2的蛋白水解降解产物，其预测质量为84 kDa，但在十二烷基硫酸钠凝胶电泳中异常运行，表观质量为110 kDa。为了促进蛋白质产物的纯化，从将六组氨酸前导序列融合到氨基末端的载体中过表达REC2基因，从而可以在固定的金属亲和柱上分离REC2蛋白。纯化的蛋白质表现出ATP依赖性的DNA复性和DNA依赖性的ATP酶活性，这是从马氏假单胞菌的细胞提取物中纯化的蛋白质的反应特征。在通过相同DNA链之间的非Watson-Crick键测量识别的测定中建立了同源配对活性。发现约50bp的大小阈值决定线性双链体分子和同源单链环之间的配对。当双链体的一条链由RNA组成时，可有效地催化与大小在阈值以下的双链体DNA形成联合分子。当存在少至三个RNA残基时，带有发夹帽的线性双链体分子也形成关节分子。

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