1887

Abstract

Vfr of is 91% similar to the cAMP receptor protein (CRP) of . Based on the high degree of sequence homology between the two proteins, the question arose whether Vfr had a global regulatory effect on gene expression for as CRP did for . This report provides two-dimensional polyacrylamide gel electrophoretic evidence that Vfr is a global regulator of gene expression in . In a :: null mutant, at least 43 protein spots were absent or decreased when compared to the proteome pattern of the parent strain. In contrast, 17 protein spots were absent or decreased in the parent strain when compared to the :: mutant. Thus, a mutation in affected production of at least 60 proteins in . In addition, the question whether Vfr and CRP shared similar mechanistic characteristics was addressed. To ascertain whether Vfr, like CRP, can bind cAMP, Vfr and CRP were purified to homogeneity and their apparent dissociation constants ( ) for binding to cAMP were determined. The values were 16 μM for Vfr and 04 μM for CRP, suggesting that these proteins have a similar affinity for cAMP. Previously the authors had demonstrated that Vfr could complement a mutation and modulate catabolite repression in . This study presents evidence that Vfr binds to the promoter and that this binding requires the presence of cAMP. Finally, the possible involvement of Vfr in catabolite repression control in was investigated. It was found that succinate repressed production of mannitol dehydrogenase, glucose-6-phosphate dehydrogenase, amidase and urocanase both in the parent and in two null mutants. This implied that catabolite repression control was not affected by the null mutation. In support of this, the cloned gene failed to complement a mutation in the gene. Thus, although Vfr is structurally similar to CRP, and is a global regulator of gene expression in , Vfr is not required for catabolite repression control in this bacterium.

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2002-05-01
2020-09-21
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