1887

Abstract

Synthesis of the [NiFe] hydrogenases 1 and 2 of is induced in response to anaerobiosis and is repressed when nitrate is present in the growth medium. The hydrogenase 1 and hydrogenase 2 enzymes are encoded by the polycistronic and operons, respectively. Primer extension analysis was used to determine the initiation site of transcription of both operons. This permitted the construction of single-copy operon fusions, which were used to examine the transcriptional regulation of the two operons. Expression of both was induced by anaerobiosis and repressed by nitrate, which is in complete accord with earlier biochemical studies. Anaerobic induction of the operon was only partially dependent on the FNR protein and, surprisingly, was enhanced by an mutation. This latter result indicated that ArcA suppresses anaerobic expression and that a further factor, which remains to be identified, is involved in controlling anaerobic induction of operon expression. Nitrate repression of expression was mediated by the NarL/NarX and NarP/NarQ two-component regulatory systems. Remarkably, a mutant lacked anaerobic induction of expression, even in the absence of added nitrate. Anaerobic induction of expression was dependent on the ArcA and AppY regulators, which confirms earlier observations by other authors. Nitrate repression of the operon was mediated by both NarL and NarP. Taken together, these data indicate that although the and operons share common regulators, there are important differences in the control of expression of the individual operons.

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1999-10-01
2024-03-29
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