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嗜热嗜酸硫化叶菌DNA依赖性RNA聚合酶大亚基A、B和C编码基因的组织与核苷酸序列

Organization and nucleotide sequence of the genes encoding the large subunits A, B and C of the DNA-dependent RNA polymerase of the archaebacterium Sulfolobus acidocaldarius.

作者信息

Pühler G, Lottspeich F, Zillig W

机构信息

Max-Planck-Institut für Biochemie, Martinsried, FRG.

出版信息

Nucleic Acids Res. 1989 Jun 26;17(12):4517-34. doi: 10.1093/nar/17.12.4517.

Abstract

The genes for the three large subunits A, B and C, of the DNA-dependent RNA polymerase of the archaebacterium Sulfolobus acidocaldarius DSM 639, were identified and characterized. The three genes follow each other immediately in the order B-A-C, which corresponds to that found in the rpoBC operon of the Escherichia coli genome. The transcription products formed in vivo were studied by Northern analysis and the start-points were determined by S1-nuclease mapping and primer directed extension analysis. The three RNA polymerase subunit genes were co-transcribed together with an open reading frame (ORF) of 88 amino acid residues length situated immediately upstream of the B gene and two ORFs of 104 and 130 amino acid residues following the C gene (together 8500 nucleotides). The following ORF, encoding a protein of 118 amino acids homologous to the ribosomal protein S12 of E. coli, was weakly transcribed with the large co-transcript and strongly from an own promoter. The derived amino acid sequence of the B-subunit was found to be homologous to the B- (second largest) subunits of the eukaryotic nuclear polymerases I, II and III and to the eubacterial beta-subunit. The combined A + C-subunits correspond to the A- (largest) subunits of the eukaryotic RNA polymerases I, II and III and to the eubacterial beta'-subunit. The amino acid sequence similarity of the Sulfolobus subunits to the eukaryotic components is clearly higher than to the E. coli subunit.

摘要

嗜热栖热菌DSM 639依赖DNA的RNA聚合酶的三个大亚基A、B和C的基因被鉴定和表征。这三个基因按B - A - C的顺序紧邻排列,这与在大肠杆菌基因组的rpoBC操纵子中发现的顺序一致。通过Northern分析研究了体内形成的转录产物,并通过S1核酸酶图谱分析和引物定向延伸分析确定了起始点。这三个RNA聚合酶亚基基因与位于B基因紧上游的一个长度为88个氨基酸残基的开放阅读框(ORF)以及C基因之后的两个长度分别为104和130个氨基酸残基的ORF(共8500个核苷酸)一起被共转录。接下来的ORF编码一种与大肠杆菌核糖体蛋白S12同源的118个氨基酸的蛋白质,它与大的共转录本一起被弱转录,并从自身启动子被强转录。发现B亚基的推导氨基酸序列与真核细胞核聚合酶I、II和III的B(第二大)亚基以及真细菌的β亚基同源。组合的A + C亚基对应于真核RNA聚合酶I、II和III的A(最大)亚基以及真细菌的β'亚基。嗜热栖热菌亚基与真核组分的氨基酸序列相似性明显高于与大肠杆菌亚基的相似性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4bcd/318011/4c094f7237ad/nar00129-0107-a.jpg

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