Transcriptional analysis of the vanC cluster from Enterococcus gallinarum strains with constitutive and inducible vancomycin resistance.
The vanC glycopeptide resistance gene cluster encodes enzymes required for synthesis of peptidoglycan precursors ending in D-Ala-D-Ser. Enterococcus gallinarum BM4174 and SC1 are constitutively and inducibly resistant to vancomycin, respectively. Analysis of peptidoglycan precursors in both strains indicated that UDP-MurNAc-tetrapeptide and UDP-MurNAc-pentapeptide[D-Ser] were synthesized in E. gallinarum SC1 only in the presence of vancomycin (4 microg/ml), whereas the "resistance" precursors accumulated in the cytoplasm of BM4174 cells under both inducing and noninducing conditions. Northern hybridization and reverse transcription-PCR experiments revealed that all the genes from the cluster, vanC-1, vanXY(C), vanT, vanR(C), and vanS(C), were transcribed from a single promoter. In the inducible SC1 isolate, transcriptional regulation appeared to be responsible for inducible expression of resistance. Promoter mapping in E. gallinarum BM4174 revealed that the transcriptional start site was located 30 nucleotides upstream from vanC-1 and that the -10 promoter consensus sequence had high identity with that of the vanA cluster. Comparison of the deduced sequence of the vanS(C) genes from isolates with constitutive and inducible resistance revealed several amino acid substitutions located in the X box (R200L) and in the region between the F and G2 boxes (D312N, D312A, and G320S) of the putative sensor kinase proteins from isolates with constitutive resistance.
Panesso
Diana
D
Bacterial Molecular Genetics Unit, Centro de Investigaciones, Universidad El Bosque, Bogotá, Columbia.
Abadía-Patiño
Lorena
L
Vanegas
Natasha
N
Reynolds
Peter E
PE
Courvalin
Patrice
P
Arias
Cesar A
CA
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D016983:Enterococcus / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D005810:Multigene Family; D010453:Peptide Synthases / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic; D020713:Vancomycin Resistance
D001426:Bacterial Proteins; D010453:Peptide Synthases; C108395:VanC protein, Enterococcus
Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7
271968
Antimicrob Agents Chemother. 1999 Sep;43(9):2161-4
10471558
Gene. 1992 Mar 1;112(1):53-8
1551598
J Bacteriol. 1992 Apr;174(8):2582-91
1556077
FEMS Microbiol Lett. 1992 Jul 1;73(1-2):195-200
1521770
Mol Microbiol. 1999 Oct;34(2):341-9
10564477
Antimicrob Agents Chemother. 2000 Jun;44(6):1660-6
10817725
J Bacteriol. 2000 Nov;182(21):6228-32
11029446
Antimicrob Agents Chemother. 2000 Nov;44(11):3224-8
11036060
Mol Microbiol. 1997 Jul;25(1):93-105
11902729
J Bacteriol. 2002 Dec;184(23):6457-64
12426332
Mol Microbiol. 2003 Nov;50(3):931-48
14617152
Mol Microbiol. 2003 Nov;50(3):1069-83
14617162
Biochimie. 1972;54(10):1291-301
4347536
Annu Rev Genet. 1992;26:71-112
1482126
J Bacteriol. 1994 Apr;176(8):2398-405
8157610
Biochem J. 1994 Jul 1;301 ( Pt 1):5-8
8037690
Nucleic Acids Res. 1994 Nov 11;22(22):4673-80
7984417
Antimicrob Agents Chemother. 1994 Sep;38(9):1899-903
7810996
Mol Microbiol. 1994 Sep;13(6):1065-70
7854121
Antimicrob Agents Chemother. 1995 Jul;39(7):1480-4
7492089
J Bacteriol. 1996 Mar;178(5):1302-9
8631706
Trends Microbiol. 1996 Oct;4(10):401-7
8899966
J Bacteriol. 1997 Jan;179(1):97-106
8981985
Antimicrob Agents Chemother. 1997 Sep;41(9):2016-8
9303405
J Bacteriol. 1998 Sep;180(17):4538-46
9721293
Proc Natl Acad Sci U S A. 1998 Sep 29;95(20):11951-6
9751771
Mol Microbiol. 1999 Apr;32(1):17-28
10216856
J Bacteriol. 1999 Jun;181(12):3644-8
10368136
Eur J Clin Microbiol Infect Dis. 1989 Nov;8(11):943-50
2532132