Characterization of the CpxRA regulon in Haemophilus ducreyi.
The Haemophilus ducreyi 35000HP genome encodes a homolog of the CpxRA two-component cell envelope stress response system originally characterized in Escherichia coli. CpxR, the cytoplasmic response regulator, was shown previously to be involved in repression of the expression of the lspB-lspA2 operon (M. Labandeira-Rey, J. R. Mock, and E. J. Hansen, Infect. Immun. 77:3402-3411, 2009). In the present study, the H. ducreyi CpxR and CpxA proteins were shown to closely resemble those of other well-studied bacterial species. A cpxA deletion mutant and a CpxR-overexpressing strain were used to explore the extent of the CpxRA regulon. DNA microarray and real-time reverse transcriptase (RT) PCR analyses indicated several potential regulatory targets for the H. ducreyi CpxRA two-component regulatory system. Electrophoretic mobility shift assays (EMSAs) were used to prove that H. ducreyi CpxR interacted with the promoter regions of genes encoding both known and putative virulence factors of H. ducreyi, including the lspB-lspA2 operon, the flp operon, and dsrA. Interestingly, the use of EMSAs also indicated that H. ducreyi CpxR did not bind to the promoter regions of several genes predicted to encode factors involved in the cell envelope stress response. Taken together, these data suggest that the CpxRA system in H. ducreyi, in contrast to that in E. coli, may be involved primarily in controlling expression of genes not involved in the cell envelope stress response.
Labandeira-Rey
Maria
M
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9048, USA.
Brautigam
Chad A
CA
Hansen
Eric J
EJ
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D015964:Gene Expression Regulation, Bacterial; D006191:Haemophilus ducreyi / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D006801:Humans; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018085:Regulon; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D037521:Virulence Factors; C085504:CpxR protein, Bacteria; D011494:Protein Kinases; C092472:CpxA protein, bacteria
Infect Immun. 2003 Dec;71(12):7178-82
14638812
J Biol Chem. 1997 Jan 17;272(3):1910-9
8999880
Infect Immun. 2004 Nov;72(11):6271-8
15501753
Cell. 2006 Sep 8;126(5):929-40
16959572
J Bacteriol. 1996 Jan;178(1):240-7
8550424
Gene. 1989 Apr 15;77(1):61-8
2744488
J Infect Dis. 1999 Jun;179(6):1423-32
10228064
Infect Immun. 1996 Jun;64(6):1950-5
8675292
J Infect Dis. 1994 May;169(5):1146-50
8169411
J Biol Chem. 2006 Nov 3;281(44):33115-26
16954205
Infect Immun. 2009 Aug;77(8):3402-11
19451237
Plasmid. 1994 Sep;32(2):228-32
7846148
Infect Immun. 1995 Apr;63(4):1241-5
7890379
Infect Immun. 2000 Mar;68(3):1608-19
10678980
Amino Acids. 2009 Sep;37(3):459-66
19241119
Microbiology (Reading). 2009 Jul;155(Pt 7):2288-2295
19389768
EMBO J. 2005 Dec 21;24(24):4247-59
16319927
FEMS Microbiol Rev. 2001 May;25(3):309-33
11348687
MMWR CDC Surveill Summ. 1992 May 29;41(3):57-61
1635549
Sex Transm Infect. 2003 Feb;79(1):68-71
12576620
J Biol Chem. 2002 Jul 19;277(29):26652-61
11953442
J Bacteriol. 2009 Mar;191(6):1798-815
19103922
J Bacteriol. 2005 Aug;187(16):5723-31
16077119
Infect Immun. 2007 Sep;75(9):4386-99
17620356
Infect Immun. 2003 Oct;71(10):5994-6003
14500520
Adv Exp Med Biol. 2008;631:80-110
18792683
Infect Immun. 2000 Nov;68(11):6441-8
11035757
J Infect Dis. 2008 Jun 1;197(11):1531-6
18422457
Infect Immun. 2004 Aug;72(8):4528-33
15271912
Mol Microbiol. 2005 Jun;56(5):1119-28
15882407
J Bacteriol. 1997 Mar;179(5):1764-73
9045839
FEMS Microbiol Lett. 1999 Jul 1;176(1):111-6
10418137
Cell. 2009 Oct 16;139(2):325-36
19800110
Adv Exp Med Biol. 2008;631:22-39
18792680
J Infect Dis. 2009 Aug 1;200(3):409-16
19552526
J Infect Dis. 2009 Mar 1;199(5):684-92
19199547
Infect Immun. 2005 Apr;73(4):2387-99
15784585
Curr Opin Microbiol. 2010 Apr;13(2):204-9
20117041
J Bacteriol. 1997 Dec;179(24):7724-33
9401031
J Infect Dis. 2009 Jun 1;199(11):1671-9
19432549
Clin Microbiol Rev. 1995 Jul;8(3):357-75
7553570
Infect Immun. 2002 Apr;70(4):1667-76
11895928
J Bacteriol. 2008 Apr;190(7):2314-22
18223085
Genes Dev. 1995 Feb 15;9(4):387-98
7883164
Infect Immun. 2003 May;71(5):2478-86
12704119
J Bacteriol. 2007 Aug;189(15):5574-81
17545286
J Bacteriol. 2001 Dec;183(24):7213-23
11717281
J Bacteriol. 1998 Nov;180(22):6013-22
9811662
J Bacteriol. 1996 Feb;178(3):808-16
8550517
Infect Immun. 2004 Apr;72(4):1874-84
15039306
Infect Immun. 2002 Jun;70(6):2965-75
12010986
J Infect Dis. 1998 Dec;178(6):1795-8
9815237
Infect Immun. 2001 Mar;69(3):1488-91
11179317
N Engl J Med. 1997 Apr 10;336(15):1072-8
9091805
Mol Microbiol. 1995 Dec;18(5):821-30
8825086
Infect Immun. 2006 May;74(5):2651-8
16622201
J Infect Dis. 2000 Mar;181(3):1049-54
10720530
Sex Transm Infect. 1999 Feb;75(1):3-17
10448335
Mol Cell. 2004 Nov 19;16(4):537-47
15546614
Annu Rev Biochem. 2000;69:183-215
10966457
Proteomics. 2007 Sep;7(17):3131-42
17676659
J Mol Biol. 1990 Oct 5;215(3):403-10
2231712
Res Microbiol. 2006 May;157(4):306-14
16487683
Infect Immun. 2006 Feb;74(2):1394-7
16428791
Microbes Infect. 2002 Sep;4(11):1141-8
12361914
J Bacteriol. 2000 Mar;182(5):1423-6
10671468
Curr Opin Infect Dis. 2010 Feb;23(1):64-9
19918177
Nucleic Acids Res. 2008 Apr;36(7):2295-300
18287115
Antimicrob Agents Chemother. 1989 Sep;33(9):1627-30
2684012