Abstract
The Streptococcus salivarius 57.I ure cluster was organized as an operon, beginning with ureI, followed by ureABC (structural genes) and ureEFGD (accessory genes). Northern analyses revealed transcripts encompassing structural genes and transcripts containing the entire operon. A sigma70-like promoter could be mapped 5' to ureI (PureI) by primer extension analysis. The intensity of the signal increased when cells were grown at an acidic pH and was further enhanced by excess carbohydrate. To determine the function(s) of two inverted repeats located 5' to PureI, transcriptional fusions of the full-length promoter region (PureI), or a deletion derivative (PureIDelta100), and a promoterless chloramphenicol acetyltransferase (CAT) gene were constructed and integrated into the chromosome to generate strains PureICAT and PureIDelta100CAT, respectively. CAT specific activities of PureICAT were repressed at pH 7.0 and induced at pH 5.5 and by excess carbohydrate. In PureIDelta100CAT, CAT activity was 60-fold higher than in PureICAT at pH 7.0 and pH induction was nearly eliminated, indicating that expression was negatively regulated. Thus, it was concluded that PureI was the predominant, regulated promoter and that regulation was governed by a mechanism differing markedly from other known mechanisms for bacterial urease expression.
MeSH Terms
Amino Acid Sequence
Bacterial Proteins/genetics,isolation & purification
Base Sequence
DNA, Bacterial
Enzyme Precursors/genetics,metabolism
Gene Expression Regulation, Bacterial
Gene Expression Regulation, Enzymologic
Membrane Transport Proteins
Molecular Sequence Data
Multigene Family
Operon
Sequence Analysis, DNA
Streptococcus/enzymology,genetics
Transcription, Genetic
Urease/genetics
Chemicals
Bacterial Proteins
DNA, Bacterial
Enzyme Precursors
Membrane Transport Proteins
UreI protein, Helicobacter pylori
Urease
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Chen Y Y
Center for Oral Biology, School of Medicine and Dentistry, University of Rochester, Rochester, New York 14642, USA.
Weaver C A
Mendelsohn D R
Burne R A
References (27)
27 references, click to expand
-
Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria.
Methods Enzymol. 1975;43:737-55
PMID: 1094240
-
Analysis of Streptococcus salivarius urease expression using continuous chemostat culture.
FEMS Microbiol Lett. 1996 Jan 15;135(2-3):223-9
PMID: 8595861
-
Urease of Klebsiella aerogenes: control of its synthesis by glutamine synthetase.
J Bacteriol. 1977 Aug;131(2):446-52
PMID: 18438
-
Characterization of two tetracycline resistance determinants in Streptococcus faecalis JH1.
J Bacteriol. 1982 May;150(2):835-43
PMID: 6802800
-
Microbial ureases: significance, regulation, and molecular characterization.
Microbiol Rev. 1989 Mar;53(1):85-108
PMID: 2651866
-
The bacteria responsible for ureolysis in artificial dental plaque.
Arch Oral Biol. 1988;33(10):727-33
PMID: 3075450
-
Regulation of gene expression and cellular localization of cloned Klebsiella aerogenes (K. pneumoniae) urease.
J Gen Microbiol. 1989 Jun;135(6):1769-76
PMID: 2693604
-
pH regulation of urease levels in Streptococcus salivarius.
J Dent Res. 1990 May;69(5):1131-7
PMID: 2110582
-
Role of the nac gene product in the nitrogen regulation of some NTR-regulated operons of Klebsiella aerogenes.
J Bacteriol. 1990 Dec;172(12):7249-55
PMID: 1979323
-
Shuttle cloning and nucleotide sequences of Helicobacter pylori genes responsible for urease activity.
J Bacteriol. 1991 Mar;173(6):1920-31
PMID: 2001995
-
Construction and properties of a family of pACYC184-derived cloning vectors compatible with pBR322 and its derivatives.
Gene. 1991 Jun 15;102(1):75-8
PMID: 1840539
-
Genetic manipulation of pathogenic streptococci.
Methods Enzymol. 1991;204:556-86
PMID: 1658571
-
Co-ordinate expression of the two threonyl-tRNA synthetase genes in Bacillus subtilis: control by transcriptional antitermination involving a conserved regulatory sequence.
EMBO J. 1992 Aug;11(8):3117-27
PMID: 1379177
-
Novel streptococcal-integration shuttle vectors for gene cloning and inactivation.
Gene. 1992 Oct 12;120(1):105-10
PMID: 1327968
-
Processes involved in the regulation of urease levels in Streptococcus salivarius by pH.
Oral Microbiol Immunol. 1992 Jun;7(3):159-64
PMID: 1408352
-
Molecular, genetic, and functional analysis of the basic replicon of pVA380-1, a plasmid of oral streptococcal origin.
Plasmid. 1992 Sep;28(2):130-45
PMID: 1409970
-
Proteus mirabilis urease: transcriptional regulation by UreR.
J Bacteriol. 1993 Jan;175(2):465-73
PMID: 7678244
-
Streptococcus mutans fructosyltransferase (ftf) and glucosyltransferase (gtfBC) operon fusion strains in continuous culture.
Infect Immun. 1993 Apr;61(4):1259-67
PMID: 8454329
-
Urease activity in Streptococcus salivarius at low pH.
Arch Oral Biol. 1993 Jun;38(6):507-16
PMID: 8343073
-
Cloning, sequencing, and expression of thermophilic Bacillus sp. strain TB-90 urease gene complex in Escherichia coli.
J Bacteriol. 1994 Jan;176(2):432-42
PMID: 8288539
-
Bacterial ureases: structure, regulation of expression and role in pathogenesis.
Mol Microbiol. 1993 Sep;9(5):907-13
PMID: 7934918
-
The Alcaligenes eutrophus protein HoxN mediates nickel transport in Escherichia coli.
J Bacteriol. 1995 Apr;177(7):1840-3
PMID: 7896709
-
Helicobacter pylori requires an acidic environment to survive in the presence of urea.
Infect Immun. 1995 May;63(5):1669-73
PMID: 7729871
-
Helicobacter pylori nickel-transport gene nixA: synthesis of catalytically active urease in Escherichia coli independent of growth conditions.
Mol Microbiol. 1995 Apr;16(1):97-109
PMID: 7651142
-
Molecular biology of microbial ureases.
Microbiol Rev. 1995 Sep;59(3):451-80
PMID: 7565414
-
Streptococcus salivarius urease: genetic and biochemical characterization and expression in a dental plaque streptococcus.
Infect Immun. 1996 Feb;64(2):585-92
PMID: 8550211
-
Transformation of Streptococcus sanguis Challis by plasmid deoxyribonucleic acid from Streptococcus faecalis.
J Bacteriol. 1976 Oct;128(1):347-55
PMID: 824275