1887

Abstract

Two unrelated protein families catalyse the oxidative decarboxylation of 2-oxoacids, i.e. the 2-oxoacid dehydrogenase complexes (OADHCs) and the 2-oxoacid ferredoxin oxidoreductases (OAFORs). In halophilic archaea, OAFORs were found to be responsible for decarboxylation of pyruvate and 2-oxoglutarate. Nevertheless, two gene clusters encoding OADHCs were found previously in , but their biological function remained obscure. Here a third gene cluster of is presented. To characterize the function, the genes encoding the E1 subunit were inactivated in all three gene clusters by in-frame deletions. Under aerobic conditions none of the three mutants showed any phenotypic difference from the wild-type in various media. However, growth yields of two mutants were considerably lower than that of wild-type under nitrate-respirative conditions in complex medium. Northern blot analyses revealed (1) that polycistronic transcripts are formed and all three gene clusters are operons and (2) that transcription of all three operons is induced under anaerobic conditions compared to aerobic conditions. Taken together, the three enzymes do not fulfil one of the ‘usual’ aerobic functions of typical OADHCs, but decarboxylate an as-yet-unidentified novel substrate under anaerobic conditions. A survey of all 28 fully sequenced archaeal genomes revealed that nearly all archaea contain several OAFORs (three to four on average), suggesting that this protein family was already present in their last common ancestor. In contrast, only nine archaea encode one or two OADHCs, indicating that this protein family entered archaea by lateral transfer of the cognate genes from bacteria. This view is underscored by a phylogenetic tree of 33 archaeal and bacterial OADHCs.

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2007-10-01
2019-11-17
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Primers used for mutant construction and expression analysis [ PDF] (22 kb) Occurrence of the genes encoding subunits of OADHCs and of OAFORs in the 28 fully sequenced genome of archaea [ PDF] (14 kb)

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Primers used for mutant construction and expression analysis [ PDF] (22 kb) Occurrence of the genes encoding subunits of OADHCs and of OAFORs in the 28 fully sequenced genome of archaea [ PDF] (14 kb)

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