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Analysis of the CO dehydrogenase/acetyl-coenzyme A synthase operon of Methanosarcina thermophila.

机译:嗜热甲烷菌的CO脱氢酶/乙酰辅酶A合酶操纵子的分析。

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摘要

The cdhABC genes encoding the respective alpha, epsilon, and beta subunits of the five-subunit (alpha, beta, gamma, delta, and epsilon) CO dehydrogenase/acetyl-coenzyme synthase (CODH/ACS) complex from Methanosarcina thermophila were cloned and sequenced. Northern (RNA) blot analyses indicated that the cdh genes encoding the five subunits and an open reading frame (ORF1) with unknown function are cotranscribed during growth on acetate. Northern blot and primer extension analyses suggested that mRNA processing and multiple promoters may be involved in cdh transcript synthesis. The putative CdhA (alpha subunit) and CdhB (epsilon subunit) proteins each have 40% identity to CdhA and CdhB of the CODH/ACS complex from Methanosaeta soehngenii. The cdhC gene encodes the beta subunit (CdhC) of the CODH/ACS complex from M. thermophila. The N-terminal 397 amino acids of CdhC are 42% identical to the C-terminal half of the alpha subunit of CODH/ACS from the acetogenic anaerobe Clostridium thermoaceticum. Sequence analysis suggested potential structures and functions for the previously uncharacterized beta subunit from M. thermophila. The deduced protein sequence of ORF1, located between the cdhC and cdhD genes, has 29% identity to NifH2 from Methanobacterium ivanovii.
机译:克隆并编码了五个嗜热甲烷单胞菌的五个亚基(α,β,γ,δ和ε)各自的α,ε和β亚基的cdhABC基因CO脱氢酶/乙酰辅酶合酶(CODH / ACS)复合物并进行了测序。 Northern(RNA)印迹分析表明,编码五个亚基的cdh基因和功能未知的开放阅读框(ORF1)在醋酸盐上生长期间共转录。 Northern印迹和引物延伸分析表明,mdh加工和多个启动子可能参与cdh转录物的合成。推定的CdhA(α亚基)和CdhB(ε亚基)蛋白各自与Methanosaeta soehngenii的CODH / ACS复合物的CdhA和CdhB具有40%的同一性。 cdhC基因编码嗜热毁丝霉CODH / ACS复合物的β亚基(CdhC)。 CdhC的N末端397个氨基酸与产热厌氧性产热梭状芽胞杆菌CODH / ACS的α亚基的C末端一半具有42%的同一性。序列分析表明嗜热毁丝霉菌先前未鉴定的β亚基的潜在结构和功能。推导的蛋白质序列ORF1位于cdhC和cdhD基因之间,与伊万诺瓦氏甲烷菌NifH2具有29%的同一性。

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