Identification and characterization of cyclic diguanylate signaling systems controlling rugosity in Vibrio cholerae.
Vibrio cholerae, the causative agent of the disease cholera, can generate rugose variants that have an increased capacity to form biofilms. Rugosity and biofilm formation are critical for the environmental survival and transmission of the pathogen, and these processes are controlled by cyclic diguanylate (c-di-GMP) signaling systems. c-di-GMP is produced by diguanylate cyclases (DGCs) and degraded by phosphodiesterases (PDEs). Proteins that contain GGDEF domains act as DGCs, whereas proteins that contain EAL or HD-GYP domains act as PDEs. In the V. cholerae genome there are 62 genes that are predicted to encode proteins capable of modulating the cellular c-di-GMP concentration. We previously identified two DGCs, VpvC and CdgA, that can control the switch between smooth and rugose. To identify other c-di-GMP signaling proteins involved in rugosity, we generated in-frame deletion mutants of all genes predicted to encode proteins with GGDEF and EAL domains and then searched for mutants with altered rugosity. In this study, we identified two new genes, cdgG and cdgH, involved in rugosity control. We determined that CdgH acts as a DGC and positively regulates rugosity, whereas CdgG does not have DGC activity and negatively regulates rugosity. In addition, epistasis analysis with CdgG, CdgH, and other DGCs and PDEs controlling rugosity revealed that CdgG and CdgH act in parallel with previously identified c-di-GMP signaling proteins to control rugosity in V. cholerae. We also determined that PilZ domain-containing c-di-GMP binding proteins contribute minimally to rugosity, indicating that there are additional c-di-GMP binding proteins controlling rugosity in V. cholerae.
Beyhan
Sinem
S
Department of Microbiology and Environmental Toxicology, University of California, Santa Cruz, Santa Cruz, CA 95064, USA.
Odell
Lindsay S
LS
Yildiz
Fitnat H
FH
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018441:Biofilms / Q000254:growth & development; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D019761:Phosphorus-Oxygen Lyases / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014734:Vibrio cholerae / Q000235:genetics / Q000378:metabolism* / Q000502:physiology
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C062025:bis(3',5')-cyclic diguanylic acid; D019761:Phosphorus-Oxygen Lyases; C060720:diguanylate cyclase; D006152:Cyclic GMP
Mol Microbiol. 2006 May;60(4):1026-43
16677312
J Infect Dis. 1996 Dec;174(6):1364-8
8940236
J Bacteriol. 2007 Mar;189(6):2319-30
17220218
Appl Environ Microbiol. 1998 Oct;64(10):3648-55
9758780
J Bacteriol. 2003 Feb;185(4):1384-90
12562809
J Biol Chem. 2007 Apr 27;282(17):12860-70
17307739
Proc Natl Acad Sci U S A. 2004 Dec 7;101(49):17084-9
15569936
J Biol Chem. 2005 Sep 2;280(35):30829-37
15994307
Clin Microbiol Rev. 2002 Oct;15(4):757-70
12364378
Mol Microbiol. 2001 Jan;39(2):223-35
11136445
J Bacteriol. 2001 Mar;183(5):1716-26
11160103
Mol Microbiol. 2007 Aug;65(4):876-95
17645452
J Bacteriol. 2004 Mar;186(5):1574-8
14973043
J Biol Chem. 2006 Oct 20;281(42):32015-24
16923812
Mol Microbiol. 2004 Apr;52(2):573-87
15066042
Appl Environ Microbiol. 2007 Nov;73(22):7482-7
17921282
J Bacteriol. 2006 Feb;188(3):1049-59
16428409
J Bacteriol. 2007 Jan;189(2):388-402
17071756
Infect Immun. 1999 Mar;67(3):1393-404
10024587
Bioinformatics. 2006 Jan 1;22(1):3-6
16249258
J Bacteriol. 2004 Aug;186(15):4864-74
15262923
J Bacteriol. 1990 Nov;172(11):6557-67
2172216
Mol Microbiol. 2007 Feb;63(4):995-1007
17233827
Microbiology (Reading). 2000 Oct;146 ( Pt 10):2395-2407
11021916
J Biol Chem. 2006 Oct 13;281(41):30310-4
16920715
Dev Cell. 2003 Oct;5(4):647-56
14536065
J Bacteriol. 2006 Apr;188(7):2681-91
16547056
Microbiol Mol Biol Rev. 1998 Dec;62(4):1301-14
9841673
FEMS Microbiol Lett. 2001 Sep 11;203(1):11-21
11557134
Mol Microbiol. 2008 Jul;69(2):376-89
18485075
Nature. 1987 Jan 15-21;325(6101):279-81
18990795
Infect Immun. 2008 Apr;76(4):1617-27
18227161
Mol Microbiol. 2006 Apr;60(2):331-48
16573684
J Bacteriol. 2005 Jul;187(14):4774-81
15995192
Infect Immun. 2005 Sep;73(9):5873-82
16113306
Mol Microbiol. 2003 Oct;50(1):101-4
14507367
Appl Environ Microbiol. 2006 Apr;72(4):2849-55
16597991
Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):4028-33
10097157
J Bacteriol. 2005 Nov;187(21):7434-43
16237027
FEMS Microbiol Lett. 2003 Oct 10;227(1):113-9
14568156
J Bacteriol. 2007 Nov;189(22):8165-78
17586641
J Bacteriol. 2007 Feb;189(3):717-29
17122338
J Bacteriol. 2006 May;188(10):3600-13
16672614
Genes Dev. 2006 Sep 15;20(18):2605-17
16980588
J Bacteriol. 2005 Mar;187(5):1792-8
15716451
Mol Microbiol. 2002 Jul;45(2):471-83
12123457
Science. 2008 Jul 18;321(5887):411-3
18635805
Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4112-7
17360486
Mol Microbiol. 2004 Jul;53(2):497-515
15228530
Mol Microbiol. 2005 Jul;57(2):420-33
15978075
Microbiology (Reading). 2007 Apr;153(Pt 4):980-994
17379708
Microbiol Immunol. 2007;51(4):369-79
17446676
Methods Enzymol. 1994;235:386-405
8057911
Gene. 1991 Dec 20;109(1):167-8
1661697
Mol Microbiol. 2007 Sep;65(6):1474-84
17824927
Proc Natl Acad Sci U S A. 2006 Apr 25;103(17):6712-7
16611728
Mol Microbiol. 2004 Aug;53(3):857-69
15255898
Curr Opin Microbiol. 2006 Apr;9(2):218-28
16530465