Araki H, Jearnpipatkul A, Tatsumi H, Sakurai T, Ushio K, Muta T, Oshima Y
J Mol Biol. 1985 Mar 20;182(2):191-203. doi: 10.1016/0022-2836(85)90338-9.
The nucleotide sequence of a 6251 base-pair plasmid, pSR1, harbored in an osmophilic haploid yeast, Zygosaccharomyces rouxii (formerly Saccharomyces rouxii), was determined. No homology was detected between the sequences of pSR1 and 2-micron DNA of Saccharomyces cerevisiae. pSR1 has a pair of inverted repeats consisting of completely homologous 959 base-pair sequences, which separate two unique sequences 2654 base-pairs and 1679 base-pairs long. Each inverted repeat has an ARS sequence functional in both Z. rouxii and S. cerevisiae hosts. Short direct repeats or dyad symmetries were observed in the inverted repeats similar to those found close to the replication origin of 2-micron DNA. Three open reading frames, P, S and R, each able to encode a protein of molecular weight larger than 10,000, were found. Insertional inactivation of R gave rise to a defect in the intramolecular recombination at the inverted repeats, and that of S reduced the copy number of pSR1 in the S. cerevisiae host. The maintenance stability of the plasmid was also tested in the heterogeneous S. cerevisiae host, but the results of the insertional inactivation of P, S and R were ambiguous. pSR1 and 2-micron DNA were compatible in S. cerevisiae cells, but the protein factors encoded by these plasmids did not complement each other.
测定了嗜高渗单倍体酵母鲁氏接合酵母(以前称为鲁氏酵母)中携带的一个6251个碱基对的质粒pSR1的核苷酸序列。未检测到pSR1序列与酿酒酵母的2μm DNA序列之间存在同源性。pSR1有一对由完全同源的959个碱基对序列组成的反向重复序列,它们将两个分别为2654个碱基对和1679个碱基对长的独特序列分隔开。每个反向重复序列都有一个在鲁氏接合酵母和酿酒酵母宿主中均起作用的自主复制序列(ARS)。在反向重复序列中观察到短的直接重复序列或二元对称序列,类似于在2μm DNA复制起点附近发现的那些序列。发现了三个开放阅读框,P、S和R,每个都能够编码分子量大于10000的蛋白质。R的插入失活导致反向重复序列处分子内重组出现缺陷,而S的插入失活降低了pSR1在酿酒酵母宿主中的拷贝数。还在异源酿酒酵母宿主中测试了该质粒的维持稳定性,但P、S和R插入失活的结果不明确。pSR1和2μm DNA在酿酒酵母细胞中是相容的,但由这些质粒编码的蛋白质因子不能相互互补。