Perozzi G, Prakash S
Mol Cell Biol. 1986 May;6(5):1497-507. doi: 10.1128/mcb.6.5.1497-1507.1986.
The RAD7 gene of Saccharomyces cerevisiae was cloned on a 4.0-kilobase (kb) DNA fragment and shown to provide full complementation of a rad7-delta mutant strain. The nucleotide sequence of a 2.2-kb DNA fragment which contains the complete RAD7 gene was determined. Transcription of the RAD7 gene initiates at multiple sites in a region spanning positions -61 to -8 of the DNA sequence. The 1.8-kb RAD7 mRNA encodes a protein of 565 amino acids with a predicted size of 63.7 kilodaltons. The hydropathy profile of the RAD7 protein indicates a highly hydrophilic amino terminus and a very hydrophobic region toward the carboxyl terminus. A RAD7 subclone deleted for the first 99 codons complements the rad7-delta mutation, but not the rad7-delta rad23-delta double mutation, indicating that the RAD23 protein can compensate for the function that is missing in the amino-terminally deleted RAD7 protein. The RAD7 and RAD23 genes in multicopy plasmids do not complement the rad23-delta and rad7-delta mutations, respectively. These observations could mean that although the two proteins might share a common functional domain, they must also perform distinct functions. Alternatively, an interaction between the RAD7 and RAD23 proteins could also account for these observations.
酿酒酵母的RAD7基因被克隆到一个4.0千碱基(kb)的DNA片段上,并显示它能完全互补rad7-δ突变菌株。测定了包含完整RAD7基因的2.2 kb DNA片段的核苷酸序列。RAD7基因的转录起始于DNA序列中-61至-8位的多个位点。1.8 kb的RAD7 mRNA编码一个由565个氨基酸组成的蛋白质,预测大小为63.7千道尔顿。RAD7蛋白的亲水性图谱显示其氨基末端高度亲水,而羧基末端有一个非常疏水的区域。缺失前99个密码子的RAD7亚克隆能互补rad7-δ突变,但不能互补rad7-δ rad23-δ双突变,这表明RAD23蛋白可以补偿氨基末端缺失的RAD7蛋白中缺失的功能。多拷贝质粒中的RAD7和RAD23基因分别不能互补rad23-δ和rad7-δ突变。这些观察结果可能意味着,尽管这两种蛋白可能共享一个共同的功能域,但它们也必须执行不同的功能。或者,RAD7和RAD23蛋白之间的相互作用也可以解释这些观察结果。