Clp and RpfF up-regulate transcription of pelA1 gene encoding the major pectate lyase in Xanthomonas campestris pv. campestris.
Exopolysaccharide and several extracellular enzymes of Xanthomonas campestris pv. campestris (Xcc), the causative agent of black rot in crucifers, are virulence determinants. In this study, two Xcc annotated extracellular pectate lyase genes, pelA1 and pelA2, belonging to family 1 of the polysaccharide lyase, were characterized. Sequence and mutational analyses have demonstrated that pelA1 encodes the major pectate lyase, whereas pelA2 is not transcribed. Using the 5' RACE method, the pelA1 transcription initiation site was mapped at nucleotide G, 103 nt upstream of the pelA1 start codon. Promoter analysis demonstrated that polygalacturonic acid and CaCl(2) induce the expression of pelA1. Transcriptional fusion assays also indicated that Clp (cAMP receptor protein-like protein) and RpfF (an enoyl-CoA hydratase homologue that is required for the synthesis of cis-11-methyl-2-dodecenoic acid, a low molecular weight diffusible signal factor, DSF) positively regulate pelA1 transcription. Gel retardation assays showed that Clp exerts a positive control over expression of pelA1 by direct binding to the upstream Clp-binding site. In conclusion, the present research demonstrated that pelA1 codes for the major pectate lyase in Xcc strain Xc17 and that its expression is up-regulated by Clp and RpfF. This is the first study to characterize pectate lyase gene expression in Xcc.
Hsiao
Yi-Min
YM
Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung 406, Taiwan, Republic of China.
Fang
Mei-Chiung
MC
Sun
Pei-Fang
PF
Tseng
Yi-Hsiung
YH
D001426:Bacterial Proteins / Q000494:pharmacology*; D002373:Cyclic AMP Receptor Protein / Q000235:genetics / Q000502:physiology; D004746:Enoyl-CoA Hydratase / Q000235:genetics / Q000502:physiology*; D016296:Mutagenesis; D011133:Polysaccharide-Lyases / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015854:Up-Regulation / Q000187:drug effects*; D016959:Xanthomonas campestris / Q000201:enzymology*
D001426:Bacterial Proteins; D002373:Cyclic AMP Receptor Protein; D011993:Recombinant Fusion Proteins; D004746:Enoyl-CoA Hydratase; D011133:Polysaccharide-Lyases; C023911:pectate lyase