title: Library screen identifies Enterococcus faecalis CcpA, the catabolite control protein A, as an effector of Ace, a collagen adhesion protein linked to virulence.
abstract: The Enterococcus faecalis cell wall-anchored protein Ace is an important virulence factor involved in cell adhesion and infection. Expression of Ace on the cell surface is affected by many factors, including stage of growth, culture temperature, and environmental components, such as serum, urine, and collagen. However, the mechanisms that regulate or modulate Ace display are not well understood. With interest in identifying genes associated with Ace expression, we utilized a whole-cell enzyme-linked immunosorbent assay (ELISA)-based screening method to identify mutants from a transposon insertion mutant library which exhibited distinct Ace surface expression profiles. We identified a ccpA insertion mutant which showed significantly decreased levels of Ace surface expression at early growth phase versus those of wild-type OG1RF. Confirmation of the observation was achieved through flow cytometry and complementation analysis. Compared to the wild type, the E. faecalis ccpA mutant had an impaired ability to adhere to collagen when grown to early exponential phase, consistent with the lack of Ace expression in the early growth phase. As a key component of carbon catabolite regulation, CcpA has been previously reported to play a critical role in regulating expression of proteins involved in E. faecalis carbohydrate uptake and utilization. Our discovery is the first to associate CcpA with the production of a major E. faecalis virulence factor, providing new insights into the regulation of E. faecalis pathogenesis.
authors: Center for Molecular Imaging, Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases, Division of Applied Biologics, The University of Texas Health Science Center at Houston, Houston, Texas, USA.
authors: Peng
authors: P
authors: Gao
authors: Kenneth L
authors: KL
authors: Pinkston
authors: Agathe
authors: A
authors: Bourgogne
authors: Melissa R
authors: MR
authors: Cruz
authors: Danielle A
authors: DA
authors: Garsin
authors: Barbara E
authors: BE
authors: Murray
authors: Barrett R
authors: BR
authors: Harvey
mesh_terms: D001422:Bacterial Adhesion / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D013293:Enterococcus faecalis / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D015723:Gene Library; D005816:Genetic Complementation Test; D009154:Mutation; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014774:Virulence
chemical_list: C121675:Ace protein, Enterococcus; D001426:Bacterial Proteins; D002352:Carrier Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger
references: J Bacteriol. 1993 Nov;175(22):7348-55
references: 8226682
references: J Bacteriol. 2002 Sep;184(18):5174-8
references: 12193635
references: BMC Microbiol. 2009 May 18;9:95
references: 19450265
references: Nat Rev Microbiol. 2008 Aug;6(8):613-24
references: 18628769
references: J Bacteriol. 1993 Aug;175(16):5216-23
references: 8349561
references: J Bacteriol. 2011 Sep;193(17):4317-25
references: 21705589
references: Res Microbiol. 1996 Jul-Sep;147(6-7):518-22
references: 9084764
references: J Bacteriol. 1974 Feb;117(2):360-72
references: 4204433
references: FEMS Microbiol Lett. 1995 Jun 15;129(2-3):207-13
references: 7607401
references: Proc Natl Acad Sci U S A. 2008 Feb 5;105(5):1698-703
references: 18230719
references: Infect Immun. 2009 Jul;77(7):2832-9
references: 19433548
references: J Bacteriol. 2008 Apr;190(7):2340-9
references: 18223086
references: BMC Microbiol. 2011 Oct 11;11:227
references: 21989394
references: J Bacteriol. 1994 Jan;176(2):511-3
references: 8288545
references: J Biol Chem. 1999 Sep 17;274(38):26939-45
references: 10480905
references: J Bacteriol. 2010 Oct;192(20):5489-98
references: 20729365
references: Microb Pathog. 2007 Aug-Sep;43(2-3):55-66
references: 17521860
references: Plasmid. 2007 Mar;57(2):131-44
references: 16996131
references: Antimicrob Agents Chemother. 2006 Apr;50(4):1183-94
references: 16569828
references: Infect Immun. 2008 Aug;76(8):3451-63
references: 18490461
references: Plasmid. 2000 Sep;44(2):183-90
references: 10964628
references: Infect Immun. 2005 Mar;73(3):1606-12
references: 15731060
references: J Bacteriol. 2000 Oct;182(20):5799-806
references: 11004180
references: Infect Immun. 2010 Jan;78(1):241-52
references: 19841076
references: J Biol Chem. 2000 Dec 22;275(51):39837-45
references: 10991941
references: Infect Immun. 2000 Sep;68(9):5218-24
references: 10948147
references: J Bacteriol. 2004 Nov;186(21):7280-9
references: 15489440
references: Mol Microbiol. 2003 Mar;47(6):1733-47
references: 12622825
references: Mol Microbiol. 1995 Mar;15(6):1049-53
references: 7623661
references: PLoS Pathog. 2010 Jan 08;6(1):e1000716
references: 20072611
references: Infect Immun. 2006 Sep;74(9):4982-9
references: 16926389
references: Infect Immun. 2000 Sep;68(9):5210-7
references: 10948146
references: Infect Immun. 2007 Nov;75(11):5399-404
references: 17785477
references: Can J Microbiol. 1978 May;24(5):512-9
references: 418859