Probing Direct Interactions between CodY and the oppD Promoter of Lactococcus lactis
| Autor: | den Hengst, CD, Curley, P, Larsen, R, Buist, G, Nauta, A, van Sinderen, D, Kuipers, OP, Kok, J, Hengst, Chris D. den |
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| Přispěvatelé: | Groningen Biomolecular Sciences and Biotechnology, Molecular Genetics, Faculty of Science and Engineering |
| Jazyk: | angličtina |
| Rok vydání: | 2005 |
| Předmět: |
Molecular Sequence Data
DNA Footprinting Microbiology CHAIN AMINO-ACIDS BACILLUS-SUBTILIS chemistry.chemical_compound HUT OPERON Bacterial Proteins Transcriptional regulation Deoxyribonuclease I NUTRITIONAL REPRESSION Gene Regulation INTRACELLULAR POOL TRANSPORT-SYSTEM Promoter Regions Genetic Molecular Biology Gene chemistry.chemical_classification biology Base Sequence DNase-I Footprinting Lactococcus lactis Mutagenesis Promoter biology.organism_classification CONTROLLED GENE-EXPRESSION Amino acid DNA-Binding Proteins GLOBAL REGULATOR CODY PEPTIDASE MUTANTS chemistry Biochemistry BACTERIA Mutagenesis Site-Directed Guanosine Triphosphate Carrier Proteins DNA Amino Acids Branched-Chain Protein Binding |
| Zdroj: | Journal of Bacteriology, 187(2), 512-521. AMER SOC MICROBIOLOGY |
| ISSN: | 0021-9193 |
| Popis: | CodY of Lactococcus lactis MG1363 is a transcriptional regulator that represses the expression of several genes encoding proteins of the proteolytic system. These genes include pepN , pepC , opp-pepO1 , and probably prtPM , pepX , and pepDA2 , since the expression of the latter three genes relative to nitrogen availability is similar to that of the former. By means of in vitro DNA binding assays and DNase I footprinting techniques, we demonstrate that L. lactis CodY interacts directly with a region upstream of the promoter of its major target known so far, the opp system. Our results indicate that multiple molecules of CodY interact with this promoter and that the amount of bound CodY molecules is affected by the presence of branched-chain amino acids and not by GTP. Addition of these amino acids strongly affects the extent of the region protected by CodY in DNase I footprints. Random and site-directed mutagenesis of the upstream region of oppD yielded variants that were derepressed in a medium with an excess of nitrogen sources. Binding studies revealed the importance of specific bases in the promoter region required for recognition by CodY. |
| Databáze: | OpenAIRE |
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