Mémet S, Gouy M, Marck C, Sentenac A, Buhler J M
Service de Biochimie, Centre d'Etudes Nucléaires de Saclay, Gif-sur-Yvette, France.
J Biol Chem. 1988 Feb 25;263(6):2830-9.
Yeast RNA polymerases are being extensively studied at the gene level. The entire gene encoding the largest subunit of RNA polymerase A, A190, was isolated and characterized in detail. Southern hybridization and gene disruption experiments showed that the RPA190 gene is unique in the haploid yeast genome and essential for cell viability. Nuclease S1 mapping was used to identify mRNA 5' and 3' termini. RPA190 encodes a polypeptide chain of 186,270 daltons in a large uninterrupted reading frame. A dot matrix comparison of the deduced amino acid sequence of subunit A190 with Escherichia coli beta' and cognate subunits B220 and C160 from yeast RNA polymerases B and C showed a conserved pattern of homology regions (I-VI). A potential DNA-binding site (zinc-binding motif) is conserved in the N-terminal region I. Remarkably, the A190 subunit does not harbor the heptapeptide repeated sequence present in the B220 subunit. The sequence of the A190 subunit diverges from B220 and C160 by the presence of two hydrophilic domains inserted between homology regions I and II, and V and VI. From their codon usage and third base pyrimidine bias, RNA polymerase genes RPA190, RPB220, RPC160, and RPC40 fall among yeast genes expressed at an average level. The RPA190 5'-flanking region contains features present in other polymerase genes that might function in regulation.
酵母RNA聚合酶正在基因层面得到广泛研究。编码RNA聚合酶A最大亚基A190的完整基因已被分离并进行了详细表征。Southern杂交和基因破坏实验表明,RPA190基因在单倍体酵母基因组中是唯一的,对细胞活力至关重要。核酸酶S1图谱分析用于鉴定mRNA的5'和3'末端。RPA190在一个大的不间断阅读框中编码一条186,270道尔顿的多肽链。将亚基A190推导的氨基酸序列与大肠杆菌β'以及酵母RNA聚合酶B和C的同源亚基B220和C160进行点阵比较,显示出同源区域(I - VI)的保守模式。一个潜在的DNA结合位点(锌结合基序)在N端区域I中是保守的。值得注意的是,A190亚基没有B220亚基中存在的七肽重复序列。A190亚基的序列与B220和C160不同,因为在同源区域I和II以及V和VI之间插入了两个亲水区。从它们的密码子使用情况和第三位碱基嘧啶偏好来看,RNA聚合酶基因RPA190、RPB220、RPC160和RPC40属于酵母中表达水平中等的基因。RPA190的5'侧翼区域包含其他聚合酶基因中可能起调控作用的特征。