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Osmotic induction of gene osmC expression in Escherichia coli K12.
J Mol Biol. 1991 Aug 20; 220(4):959-73.JM

Abstract

osmC, an osmotically inducible gene of Escherichia coli, was physically mapped on the bacterial chromosome, cloned on multicopy plasmids, and its product, OsmC, was identified as a 14 kDa protein in maxicells. The DNA sequence of the gene and its upstream region were determined. The sequence of an osmC-phoA gene fusion confirmed the osmC reading frame. A deletion of osmC from the E. coli chromosome was constructed by gene replacement, demonstrating that it is not an essential gene. The osmCp promoter region was subcloned and a lac operon fusion transcribed under osmCp control was constructed. The expression of this operon fusion demonstrated that osmC regulation occurs at the transcriptional level. S1 nuclease protection experiments and deletion analysis identified two overlapping promoters with transcription start sites separated by ten nucleotides. All the sequences necessary for osmotic regulation of both promoters are located within a 137 base-pair DNA fragment extending from position -95 to +42 with respect to the putative osmC translation start. Two deletions were obtained that abolish the functioning of the upstream promoter. Yet, under our experimental conditions, the subsequent expression of the osmC-lacZ fusion was equivalent to that obtained from the tandem promoters. Mutations leading to constitutive expression of osmC were selected. Two independent mutations were obtained, both affected osmZ, the gene encoding the histone-like protein H1.

Authors+Show Affiliations

Centre de Recherche de Biochimie et Génétique Cellulaire du CNRS, Toulouse, France.No affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

1715407

Citation

Gutierrez, C, and J C. Devedjian. "Osmotic Induction of Gene osmC Expression in Escherichia Coli K12." Journal of Molecular Biology, vol. 220, no. 4, 1991, pp. 959-73.
Gutierrez C, Devedjian JC. Osmotic induction of gene osmC expression in Escherichia coli K12. J Mol Biol. 1991;220(4):959-73.
Gutierrez, C., & Devedjian, J. C. (1991). Osmotic induction of gene osmC expression in Escherichia coli K12. Journal of Molecular Biology, 220(4), 959-73.
Gutierrez C, Devedjian JC. Osmotic Induction of Gene osmC Expression in Escherichia Coli K12. J Mol Biol. 1991 Aug 20;220(4):959-73. PubMed PMID: 1715407.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Osmotic induction of gene osmC expression in Escherichia coli K12. AU - Gutierrez,C, AU - Devedjian,J C, PY - 1991/8/20/pubmed PY - 1991/8/20/medline PY - 1991/8/20/entrez SP - 959 EP - 73 JF - Journal of molecular biology JO - J Mol Biol VL - 220 IS - 4 N2 - osmC, an osmotically inducible gene of Escherichia coli, was physically mapped on the bacterial chromosome, cloned on multicopy plasmids, and its product, OsmC, was identified as a 14 kDa protein in maxicells. The DNA sequence of the gene and its upstream region were determined. The sequence of an osmC-phoA gene fusion confirmed the osmC reading frame. A deletion of osmC from the E. coli chromosome was constructed by gene replacement, demonstrating that it is not an essential gene. The osmCp promoter region was subcloned and a lac operon fusion transcribed under osmCp control was constructed. The expression of this operon fusion demonstrated that osmC regulation occurs at the transcriptional level. S1 nuclease protection experiments and deletion analysis identified two overlapping promoters with transcription start sites separated by ten nucleotides. All the sequences necessary for osmotic regulation of both promoters are located within a 137 base-pair DNA fragment extending from position -95 to +42 with respect to the putative osmC translation start. Two deletions were obtained that abolish the functioning of the upstream promoter. Yet, under our experimental conditions, the subsequent expression of the osmC-lacZ fusion was equivalent to that obtained from the tandem promoters. Mutations leading to constitutive expression of osmC were selected. Two independent mutations were obtained, both affected osmZ, the gene encoding the histone-like protein H1. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/1715407/Osmotic_induction_of_gene_osmC_expression_in_Escherichia_coli_K12_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0022-2836(91)90366-E DB - PRIME DP - Unbound Medicine ER -