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dc.contributor | Facultad de Ciencias Biologicas y Ambientales | es_ES |
dc.contributor.author | Santamarta, Irene | |
dc.contributor.author | Rodríguez García, Antonio | |
dc.contributor.author | Pérez Redondo, Rosario | |
dc.contributor.author | Martín Martín, Juan Francisco | |
dc.contributor.author | Liras Padín, Paloma | |
dc.contributor.other | Microbiologia | es_ES |
dc.date | 2002-03-05 | |
dc.date.accessioned | 2024-01-22T20:17:00Z | |
dc.date.available | 2024-01-22T20:17:00Z | |
dc.identifier.citation | Santamarta, I., Rodríguez-García, A., Pérez-Redondo, R., Martín, J. F., & Liras, P. (2002). CcaR is an autoregulatory protein that binds to the ccaR and cefD-cmcI promoters of the cephamycin C-clavulanic acid cluster in Streptomyces clavuligerus. Journal of Bacteriology, 184(11), 3106-3113. https://doi.org/10.1128/JB.184.11.3106-3113.2002 | es_ES |
dc.identifier.issn | 0021-9193 | |
dc.identifier.other | https://journals.asm.org/doi/10.1128/jb.184.11.3106-3113.2002 | es_ES |
dc.identifier.uri | https://hdl.handle.net/10612/17714 | |
dc.description.abstract | [EN] The putative regulatory CcaR protein, which is encoded in the β-lactam supercluster of Streptomyces clavuligerus, has been partially purified by ammonium sulfate precipitation and heparin affinity chromatography. In addition, it was expressed in Escherichia coli, purified as a His-tagged recombinant protein (rCcaR), and used to raise anti-rCcaR antibodies. The partially purified CcaR protein from S. clavuligerus was able to bind DNA fragments containing the promoter regions of the ccaR gene itself and the bidirectional cefD-cmcI promoter region. In contrast, CcaR did not bind to DNA fragments with the promoter regions of other genes of the cephamycin-clavulanic acid supercluster including lat, blp, claR, car-cyp, and the unlinked argR gene. The DNA shifts obtained with CcaR were prevented by anti-rCcaR immunoglobulin G (IgG) antibodies but not by anti-rabbit IgG antibodies. ccaR and the bidirectional cefD-cmcI promoter region were fused to the xylE reporter gene and expressed in Streptomyces lividans and S. clavuligerus. These constructs produced low catechol dioxygenase activity in the absence of CcaR; activity was increased 1.7- to 4.6-fold in cultures expressing CcaR. Amplification of the ccaR promoter region lacking its coding sequence in a high-copy-number plasmid in S. clavuligerus ATCC 27064 resulted in a reduced production of cephamycin C and clavulanic acid, by 12 to 20% and 40 to 60%, respectively, due to titration of the CcaR regulator. These findings confirm that CcaR is a positively acting autoregulatory protein able to bind to its own promoter as well as to the cefD-cmcI bidirectional promoter region | es_ES |
dc.language | eng | es_ES |
dc.publisher | American Society for Microbiology | es_ES |
dc.subject | Biotecnología | es_ES |
dc.subject.other | Cephamycin C | es_ES |
dc.subject.other | Clavulanic Acid | es_ES |
dc.subject.other | Transcriptional regulation | es_ES |
dc.subject.other | CcaR | es_ES |
dc.subject.other | Streptomyces clavuligerus | es_ES |
dc.subject.other | Autorregulation | es_ES |
dc.subject.other | Biosynthesis gene cluster | es_ES |
dc.subject.other | Secondary metabolism | es_ES |
dc.title | CcaR is an autoregulatory protein that binds to the ccaR and cefD-cmcI promoters of the cephamycin C-clavulanic acid cluster in Streptomyces clavuligerus | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.identifier.doi | 10.1128/JB.184.11.3106-3113.2002 | |
dc.description.peerreviewed | SI | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/MICYT//FD97-1419-CO2-O2/ES/ | es_ES |
dc.rights.accessRights | info:eu-repo/semantics/embargoedAccess | es_ES |
dc.identifier.essn | 1098-5530 | |
dc.journal.title | Journal of Bacteriology | es_ES |
dc.volume.number | 184 | es_ES |
dc.issue.number | 11 | es_ES |
dc.page.initial | 3106 | es_ES |
dc.page.final | 3113 | es_ES |
dc.type.hasVersion | info:eu-repo/semantics/publishedVersion | es_ES |
dc.subject.unesco | 2414.02 Fisiología Bacteriana | es_ES |
dc.subject.unesco | 2415.01 Biología Molecular de Microorganismos | es_ES |
dc.description.project | This work was supported by the Spanish Ministry of Science and Technology (FD97-1419-CO2-O2). I. Santamarta received a fellowship from the University of León. We are grateful to A. de la Fuente, F. J. Enguita, and C. de Torre for their interest and helpful discussions, to A. Jiménez for revising the manuscript, and to M. Mediavilla for technical assistance. | es_ES |
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