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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.

机译:编码古细菌Sulfolobus acidocaldarius的DNA依赖性RNA聚合酶大亚基A,B和C的基因的组织和核苷酸序列。

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摘要

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.
机译:鉴定并鉴定了古细菌Sulfolobus acidocaldarius DSM 639的DNA依赖性RNA聚合酶的三个大亚基A,B和C的基因。这三个基因以B-A-C的顺序相互跟随,这与在大肠杆菌基因组的rpoBC操纵子中发现的顺序相对应。通过Northern分析研究了体内形成的转录产物,并通过S1-核酸酶作图和引物指导的延伸分析确定了起点。将三个RNA聚合酶亚基基因与紧接B基因上游的88个氨基酸残基的开放阅读框(ORF)和紧随C基因之后的104和130个氨基酸残基的两个ORF(共8500个核苷酸)一起共转录)。编码与大肠杆菌核糖体蛋白S12同源的118个氨基酸的蛋白质的以下ORF被较大的共转录物弱转录,并强烈地自启动子转录。发现B-亚基的衍生氨基酸序列与真核核聚合酶I,II和III的B-(第二大)亚基和真细菌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,并且对应于真细菌β'-亚基。硫磺亚单位与真核生物组分的氨基酸序列相似性明显高于大肠杆菌亚单位。

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