Shpakovski G V
Laboratory of Molecular Biology, National Institute of Mental Health, Bethesda, MD 20892.
Gene. 1994 Sep 15;147(1):63-9. doi: 10.1016/0378-1119(94)90039-6.
A single-copy gene, homologous to the RPB6 gene from Saccharomyces cerevisiae, encoding a small phosphorylated subunit common to all three forms of nuclear DNA-dependent RNA polymerase was isolated from the fission yeast Schizosaccharomyces pombe. Its cDNA copy consists of an open reading frame of 142 codons and encodes an acidic protein (predicted pI 4.1) with a M(r) of 15,730. The genomic copy of Sz. pombe rpb6 contains an intron (219 nucleotides) located at codon 92, a position which does not correspond to the single intron of the S. cerevisiae gene. The sequencing of both genomic and cDNA copies of rpb6 allowed us to determine the probable positions of the start and stop of rpb6 transcription and to identify a putative TATA box. The primary structures of the Sz. pombe and S. cerevisiae Rpb6 proteins have 60.7% identity, with the same general organization: a highly acidic N-terminal region followed by a short basic region and a C terminus featuring a putative heptad Leu repeat. The C-terminal half of the sequence is particularly well conserved and, therefore, probably contains the most important functional domain. Moreover, a heterospecific complementation test showed that rpb6 from Sz. pombe fully complements a complete deletion of its S. cerevisiae homologue.
从裂殖酵母粟酒裂殖酵母中分离出一个单拷贝基因,它与酿酒酵母的RPB6基因同源,编码三种形式的核DNA依赖性RNA聚合酶共有的一个小的磷酸化亚基。其cDNA拷贝由142个密码子的开放阅读框组成,编码一种酸性蛋白(预测pI为4.1),分子量为15,730。粟酒裂殖酵母rpb6的基因组拷贝包含一个位于第92位密码子处的内含子(219个核苷酸),该位置与酿酒酵母基因的单个内含子位置不对应。rpb6的基因组和cDNA拷贝的测序使我们能够确定rpb6转录起始和终止的可能位置,并鉴定出一个推定的TATA框。粟酒裂殖酵母和酿酒酵母Rpb6蛋白的一级结构具有60.7%的同一性,具有相同的总体结构:一个高度酸性的N端区域,后面跟着一个短的碱性区域和一个以推定的七肽亮氨酸重复序列为特征的C端。该序列的C端一半特别保守,因此可能包含最重要的功能域。此外,异源互补试验表明,粟酒裂殖酵母的rpb6完全互补其酿酒酵母同源物的完全缺失。