Yamada H, Mori H, Momoi H, Nakagawa Y, Ueguchi C, Mizuno T
Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Japan.
Yeast. 1994 Jul;10(7):883-94. doi: 10.1002/yea.320100704.
We searched for fission yeast (Schizosaccharomyces pombe) proteins that preferentially bind to a synthetic curved DNA sequence, by means of a DNA-binding gel shift assay in the presence of an excess amount of a non-curved DNA sequence as a competitor. We identified such a protein in S. pombe. The protein, thus purified, has an apparent molecular weight of 42,000, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was suggested that this protein (42 K-protein) recognizes and binds to a curved DNA structure in a given nucleotide sequence, although it also binds to a non-curved DNA sequence with lower affinity. As its putative coding sequence, a 1.9-kilobase genomic DNA from S. pombe was cloned and sequenced. Sequencing of a cDNA clone also revealed the existence of an open reading frame, with no intron, encoding a 381-amino-acid protein with a calculated molecular mass, 41,597. This protein appears to be located in the nucleus. The predicted protein sequence revealed that the 42 K-protein exhibits no significant similarity to any other known proteins, except to a hypothetical protein of Caenorhabditis elegans.
我们通过DNA结合凝胶迁移试验,在存在过量非弯曲DNA序列作为竞争者的情况下,寻找优先结合合成弯曲DNA序列的裂殖酵母(粟酒裂殖酵母)蛋白。我们在粟酒裂殖酵母中鉴定出了这样一种蛋白。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳估计,纯化后的该蛋白表观分子量为42,000。有人提出,这种蛋白(42K蛋白)在特定核苷酸序列中识别并结合弯曲的DNA结构,尽管它也以较低亲和力结合非弯曲DNA序列。作为其推定的编码序列,克隆并测序了来自粟酒裂殖酵母的1.9千碱基基因组DNA。一个cDNA克隆的测序也揭示了一个无内含子的开放阅读框,编码一个381个氨基酸的蛋白,计算分子量为41,597。这种蛋白似乎位于细胞核中。预测的蛋白序列显示,42K蛋白与任何其他已知蛋白均无显著相似性,除了与秀丽隐杆线虫的一种假定蛋白相似。