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|標題:||Clp upregulates transcription of engA gene encoding a virulence factor in Xanthomonas campestris by direct binding to the upstream tandem Clp sites||作者:||Hsiao, Y.M.
|關鍵字:||Clp;transcriptional fussion assay;endoglucanase;electrophoretic;mobility shift assay;Xanthomonas campestris;amp receptor protein;pathogenicity factor production;phage phi-lf;escherichia-coli;pv campestris;pathovar campestris;activator protein;molecular analysis;rna-polymerase;dna||Project:||Febs Letters||期刊/報告no：:||Febs Letters, Volume 579, Issue 17, Page(s) 3525-3533.||摘要:||
In Xanthomonas campestris, the causative agent of black rot in crucifers, the endoglucanase level is greatly decreased in the mutant deficient in Clip, a homologue of cyclic AMP receptor protein (CRP). It is established that Clp has the same DNA binding specificity as CRP at positions 5, 6, and 7 (GTG motif) of the DNA half site. In this study, the engA transcription initiation site was determined by the 5' RACE method, and two consensus Clp-binding sites, site I and site II centered at -69.5 and -42.5, respectively, were located. Transcriptional fusion assays indicated that Clp greatly activates engA transcription. Site-directed mutagenesis indicated that position 5 of GTG motif in site II is essential for both DNA-protein complex formation in electrophoretic mobility shift assays and engA transcription in vivo. In addition, mutation at position 5 of site I drastically reduces the promoter activity, indicating that binding of Clp to site I exerts a synergistic effect on the transcription activation by site II. engA appears to be the first X. campestris gene known to be activated by Clp via a direct binding to the promoter. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V.. All rights reserved.
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