Larimer F W, Perry J R, Hardigree A A
Biology Division, Oak Ridge National Laboratory, Tennessee 37831.
J Bacteriol. 1989 Jan;171(1):230-7. doi: 10.1128/jb.171.1.230-237.1989.
The REV1 gene of Saccharomyces cerevisiae is required for normal induction of mutations by physical and chemical agents. We have determined the sequence of a 3,485-base-pair segment of DNA that complements the rev1-1 mutant. Gene disruption was used to confirm that this DNA contained the REV1 gene. The sequenced segment contains a single long open reading frame, which can encode a polypeptide of 985 amino acid residues. The REV1 transcript is 3.1 kilobase pairs in length. Frameshift mutations introduced into the open reading frame yielded a Rev-phenotype. A base substitution, encoding Gly-193 to Arg-193, was found in this open reading frame in rev1-1. Deletion mutants, lacking segments of the 5' region of REV1, had intermediate mutability relative to REV1 and rev1-1; a complete deletion exhibited lower mutability than rev1-1. REV1 is not an essential gene. An in-frame fusion of the 5' end of the REV1 open reading frame to the lacZ gene produced beta-galactosidase activity constitutively. The predicted REV1 protein is hydrophilic, with a predicted pI of 9.82. No homologies to RAD1, RAD2, RAD3, RAD7, or RAD10 proteins were noted. A 152-residue internal segment displayed 25% identity with UMUC protein.
酿酒酵母的REV1基因是物理和化学诱变剂正常诱导突变所必需的。我们已经确定了一段3485个碱基对的DNA序列,它能互补rev1 - 1突变体。通过基因敲除证实该DNA包含REV1基因。测序片段包含一个单一的长开放阅读框,可编码一个由985个氨基酸残基组成的多肽。REV1转录本长度为3.1千碱基对。引入开放阅读框的移码突变产生Rev表型。在rev1 - 1的这个开放阅读框中发现了一个碱基替换,将甘氨酸 - 193编码为精氨酸 - 193。缺失REV1 5'区域片段的缺失突变体,其变异性相对于REV1和rev1 - 1为中等;完全缺失表现出比rev1 - 1更低的变异性。REV1不是必需基因。REV1开放阅读框5'端与lacZ基因的读框内融合持续产生β - 半乳糖苷酶活性。预测的REV1蛋白是亲水性的,预测的pI为9.82。未发现与RAD1、RAD2、RAD3、RAD7或RAD10蛋白有同源性。一个152个残基的内部片段与UMUC蛋白有25%的同一性。