The PecT repressor interacts with regulatory regions of pectate lyase genes in Erwinia chrysanthemi
| Autor: | William Nasser, Arnaud Castillo, Guy Condemine, Sylvie Reverchon |
|---|---|
| Přispěvatelé: | Laboratoire de Génétique Moléculaire des Microorganismes et des Interactions Cellulaires (LGMMIC), Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Rodrigue, Agnès |
| Jazyk: | angličtina |
| Rok vydání: | 1998 |
| Předmět: |
Transcription
Genetic [SDV]Life Sciences [q-bio] Mutant Molecular Sequence Data Biophysics Repressor Virulence Biology Regulatory Sequences Nucleic Acid Biochemistry Polymerase Chain Reaction 03 medical and health sciences Bacterial Proteins Structural Biology Genetics Escherichia coli RNA Messenger Cloning Molecular Gene 030304 developmental biology Plant Diseases Polysaccharide-Lyases 0303 health sciences Molecular mass Base Sequence 030306 microbiology Dickeya chrysanthemi Plants Phenotype Molecular biology [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology Recombinant Proteins [SDV] Life Sciences [q-bio] Repressor Proteins Regulatory sequence Pectate lyase Trans-Activators [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology Plasmids Transcription Factors |
| Zdroj: | Biochim Biophys Acta Biochim Biophys Acta, 1998, 1442 (2-3), pp.148-60 |
| Popis: | Erwinia chrysanthemi is a broad host range phytopathogenic enterobacterium responsible for soft-rot disease of many plant species. The pecT gene encodes a repressor that negatively regulates the expression of virulence factors, such as pectinases, motility or exopolysaccharide synthesis. The cloned pecT gene was overexpressed using a phage T7 system. The purification of PecT involved the use of a TSK-heparin column and delivered the PecT protein that was purified to near homogeneity. The purified repressor displayed a 34 kDa apparent molecular mass. Gel-filtration experiments revealed that the PecT protein is a dimer. Band-shift assays demonstrated that the tetramer of the PecT protein could specifically bind in vitro to the regulatory regions of the pectate lyase genes with variable affinities. In addition, we demonstrated that PecT represses its own synthesis by interacting independently with two 200 bp regions, R1 and R2, located from −382 to −632 and −17 to −234, respectively, from the distal P1 promoter and from −465 to −715 and −100 to −317 from the P2 proximal promoter. We propose a model that explains the regulation exerted by PecT on its target genes and that integrates the phenotype obtained with a PecT overproducing pec-1 mutant or a pecT mutant. |
| Databáze: | OpenAIRE |
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