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Specific interactions of Erwinia chrysanthemi KdgR repressor with different operators of genes involved in pectinolysis.
J Mol Biol. 1994 Feb 18; 236(2):427-40.JM

Abstract

The Erwinia chrysanthemi kdgR gene encodes a repressor that negatively regulates the expression of genes involved in pectinolysis and in pectinase secretion. The cloned kdgR gene was overexpressed in Escherichia coli by using a phage T7 system. Overproduced repressor was purified to homogeneity by two chromatographic steps. Gel retardation and DNase I protection experiments demonstrated the specific binding of the KdgR protein to the operators of pectinase genes (pelA, pelB, pelC, pelE), to the operator of genes involved in pectin catabolism (kdgT, ogl, kduI-kdgF) and to that of the outT gene involved in pectinase secretion. These interactions involved one (pelA, pelB, kduI-kdgF, outT) or several operator sites (pelC, pelE, ogl, kdgT) that generally overlap the promoter. Despite the presence of potential KdgR binding sites (KdgR-box) in the regulatory regions of four genes involved in pectin catabolism (kdgC, kduD, pem, kdgA) and in a pectinase secretion gene outC, no DNA-repressor complex could be observed by in vitro experiments. By using a missing contact experiment on the coding strand of ogl and pelE regulatory regions, a new KdgR-binding consensus was proposed. This new consensus, constituted by two half motifs (AATGAAAACT)N(NTCGATTTCTA), is well conserved in the operators which interact in vitro with the KdgR repressor. In contrast, this repressor-recognized motif is degenerated in the other operators that cannot interact in vitro with the repressor. These results suggest the existence of different regulation mechanisms mediated by the KdgR protein for the two classes of operators.

Authors+Show Affiliations

Laboratoire de Génétique Moléculaire des Microorganismes et des Interactions Cellulaires, URA CNRS 1486, Institut National des Sciences Appliqées, Villeurbanne, France.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8107132

Citation

Nasser, W, et al. "Specific Interactions of Erwinia Chrysanthemi KdgR Repressor With Different Operators of Genes Involved in Pectinolysis." Journal of Molecular Biology, vol. 236, no. 2, 1994, pp. 427-40.
Nasser W, Reverchon S, Condemine G, et al. Specific interactions of Erwinia chrysanthemi KdgR repressor with different operators of genes involved in pectinolysis. J Mol Biol. 1994;236(2):427-40.
Nasser, W., Reverchon, S., Condemine, G., & Robert-Baudouy, J. (1994). Specific interactions of Erwinia chrysanthemi KdgR repressor with different operators of genes involved in pectinolysis. Journal of Molecular Biology, 236(2), 427-40.
Nasser W, et al. Specific Interactions of Erwinia Chrysanthemi KdgR Repressor With Different Operators of Genes Involved in Pectinolysis. J Mol Biol. 1994 Feb 18;236(2):427-40. PubMed PMID: 8107132.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Specific interactions of Erwinia chrysanthemi KdgR repressor with different operators of genes involved in pectinolysis. AU - Nasser,W, AU - Reverchon,S, AU - Condemine,G, AU - Robert-Baudouy,J, PY - 1994/2/18/pubmed PY - 1994/2/18/medline PY - 1994/2/18/entrez SP - 427 EP - 40 JF - Journal of molecular biology JO - J Mol Biol VL - 236 IS - 2 N2 - The Erwinia chrysanthemi kdgR gene encodes a repressor that negatively regulates the expression of genes involved in pectinolysis and in pectinase secretion. The cloned kdgR gene was overexpressed in Escherichia coli by using a phage T7 system. Overproduced repressor was purified to homogeneity by two chromatographic steps. Gel retardation and DNase I protection experiments demonstrated the specific binding of the KdgR protein to the operators of pectinase genes (pelA, pelB, pelC, pelE), to the operator of genes involved in pectin catabolism (kdgT, ogl, kduI-kdgF) and to that of the outT gene involved in pectinase secretion. These interactions involved one (pelA, pelB, kduI-kdgF, outT) or several operator sites (pelC, pelE, ogl, kdgT) that generally overlap the promoter. Despite the presence of potential KdgR binding sites (KdgR-box) in the regulatory regions of four genes involved in pectin catabolism (kdgC, kduD, pem, kdgA) and in a pectinase secretion gene outC, no DNA-repressor complex could be observed by in vitro experiments. By using a missing contact experiment on the coding strand of ogl and pelE regulatory regions, a new KdgR-binding consensus was proposed. This new consensus, constituted by two half motifs (AATGAAAACT)N(NTCGATTTCTA), is well conserved in the operators which interact in vitro with the KdgR repressor. In contrast, this repressor-recognized motif is degenerated in the other operators that cannot interact in vitro with the repressor. These results suggest the existence of different regulation mechanisms mediated by the KdgR protein for the two classes of operators. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/8107132/Specific_interactions_of_Erwinia_chrysanthemi_KdgR_repressor_with_different_operators_of_genes_involved_in_pectinolysis_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(84)71155-7 DB - PRIME DP - Unbound Medicine ER -