title: Functional identification of conserved residues involved in Lactobacillus rhamnosus strain GG sortase specificity and pilus biogenesis.
abstract: In Gram-positive bacteria, sortase-dependent pili mediate the adhesion of bacteria to host epithelial cells and play a pivotal role in colonization, host signaling, and biofilm formation. Lactobacillus rhamnosus strain GG, a well known probiotic bacterium, also displays on its cell surface mucus-binding pilus structures, along with other LPXTG surface proteins, which are processed by sortases upon specific recognition of a highly conserved LPXTG motif. Bioinformatic analysis of all predicted LPXTG proteins encoded by the L. rhamnosus GG genome revealed a remarkable conservation of glycine residues juxtaposed to the canonical LPXTG motif. Here, we investigated and defined the role of this so-called triple glycine (TG) motif in determining sortase specificity during the pilus assembly and anchoring. Mutagenesis of the TG motif resulted in a lack or an alteration of the L. rhamnosus GG pilus structures, indicating that the TG motif is critical in pilus assembly and that they govern the pilin-specific and housekeeping sortase specificity. This allowed us to propose a regulatory model of the L. rhamnosus GG pilus biogenesis. Remarkably, the TG motif was identified in multiple pilus gene clusters of other Gram-positive bacteria, suggesting that similar signaling mechanisms occur in other, mainly pathogenic, species.
authors.affiliation: From the Department of Veterinary Biosciences, University of Helsinki, Helsinki 00790, Finland and francois.douillard@helsinki.fi.
authors.forename: François P
authors.initials: FP
authors.lastname: Douillard
authors.affiliation: From the Department of Veterinary Biosciences, University of Helsinki, Helsinki 00790, Finland and.
authors.forename: Pia
authors.initials: P
authors.lastname: Rasinkangas
authors.affiliation: From the Department of Veterinary Biosciences, University of Helsinki, Helsinki 00790, Finland and.
authors.forename: Ingemar
authors.initials: I
authors.lastname: von Ossowski
authors.affiliation: From the Department of Veterinary Biosciences, University of Helsinki, Helsinki 00790, Finland and.
authors.forename: Justus
authors.initials: J
authors.lastname: Reunanen
authors.affiliation: From the Department of Veterinary Biosciences, University of Helsinki, Helsinki 00790, Finland and.
authors.forename: Airi
authors.initials: A
authors.lastname: Palva
authors.affiliation: From the Department of Veterinary Biosciences, University of Helsinki, Helsinki 00790, Finland and the Laboratory of Microbiology, Wageningen University, 6708 PB Wageningen, The Netherlands.
authors.forename: Willem M
authors.initials: WM
authors.lastname: de Vos
mesh_terms: D019881:Aminoacyltransferases / Q000235:genetics / Q000378:metabolism*; D001422:Bacterial Adhesion / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D036781:Fimbriae Proteins / Q000235:genetics / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000201:enzymology* / Q000648:ultrastructure; D005998:Glycine / Q000235:genetics; D052201:Lacticaseibacillus rhamnosus / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D016297:Mutagenesis, Site-Directed; D019936:Probiotics; D015398:Signal Transduction / Q000502:physiology; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D036781:Fimbriae Proteins; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases; D005998:Glycine
keywords: Bacterial Adhesion; Electron Microscopy (EM); Protein Motif; Signaling; Site-directed Mutagenesis
references.citation: Nature. 2003 May 1;423(6935):87-91
references.pmid: 12721630
references.citation: Trends Microbiol. 2008 Jan;16(1):33-40
references.pmid: 18083568
references.citation: Appl Environ Microbiol. 1989 Dec;55(12):3119-23
references.pmid: 16348073
references.citation: PLoS One. 2009 Jul 15;4(7):e6072
references.pmid: 19603075
references.citation: FASEB J. 2013 Aug;27(8):3144-54
references.pmid: 23631841
references.citation: Cell Mol Life Sci. 2009 Feb;66(4):613-35
references.pmid: 18953686
references.citation: Microbiology (Reading). 2010 Sep;156(Pt 9):2649-2659
references.pmid: 20542929
references.citation: Vet Microbiol. 2009 Jul 2;138(1-2):132-9
references.pmid: 19303725
references.citation: PLoS One. 2013 May 14;8(5):e64416
references.pmid: 23691212
references.citation: Appl Environ Microbiol. 2007 Nov;73(21):6768-75
references.pmid: 17827316
references.citation: Mol Microbiol. 2007 Oct;66(2):495-510
references.pmid: 17897374
references.citation: Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Oct;69(Pt 10):1182-5
references.pmid: 24100577
references.citation: Nat Rev Microbiol. 2010 Mar;8(3):171-84
references.pmid: 20157338
references.citation: Infect Immun. 2004 May;72(5):2710-22
references.pmid: 15102780
references.citation: Science. 2001 Jul 20;293(5529):498-506
references.pmid: 11463916
references.citation: J Biol Chem. 2009 Sep 4;284(36):24465-77
references.pmid: 19592495
references.citation: Mol Microbiol. 2007 Nov;66(4):961-74
references.pmid: 17919283
references.citation: Appl Environ Microbiol. 2010 Apr;76(7):2049-57
references.pmid: 20118368
references.citation: Mol Microbiol. 2003 Nov;50(4):1429-38
references.pmid: 14622427
references.citation: Curr Opin Biotechnol. 2005 Apr;16(2):204-11
references.pmid: 15831388
references.citation: Appl Environ Microbiol. 2012 Jan;78(1):185-93
references.pmid: 22020518
references.citation: Science. 2005 Jul 1;309(5731):105
references.pmid: 15994549
references.citation: Appl Environ Microbiol. 2013 Mar;79(6):1923-33
references.pmid: 23315726
references.citation: Biochemistry. 2009 Nov 10;48(44):10549-57
references.pmid: 19810750
references.citation: FEMS Microbiol Rev. 2006 Jul;30(4):487-513
references.pmid: 16774584
references.citation: Infect Immun. 2006 Apr;74(4):2453-6
references.pmid: 16552078
references.citation: Gene. 1997 Jun 11;192(1):155-63
references.pmid: 9224886
references.citation: Appl Environ Microbiol. 2001 Feb;67(2):608-16
references.pmid: 11157223
references.citation: ACS Nano. 2013 Apr 23;7(4):3685-97
references.pmid: 23531039
references.citation: Nat Rev Microbiol. 2006 Jul;4(7):509-19
references.pmid: 16778837
references.citation: Science. 2003 Mar 28;299(5615):2071-4
references.pmid: 12663927
references.citation: Jpn J Vet Res. 1968 Mar;16(1):31-7
references.pmid: 5304169
references.citation: J Biol Chem. 2008 Dec 26;283(52):36676-86
references.pmid: 18940793
references.citation: Infect Immun. 2011 Jul;79(7):2911-20
references.pmid: 21502588
references.citation: BMC Microbiol. 2008 Oct 01;8:165
references.pmid: 18828902
references.citation: Appl Environ Microbiol. 2010 Nov;76(21):7332-7
references.pmid: 20851967
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):13950-5
references.pmid: 16172379
references.citation: J Bacteriol. 2006 Sep;188(17):6318-25
references.pmid: 16923899
references.citation: Proc Natl Acad Sci U S A. 2002 Sep 17;99(19):12391-6
references.pmid: 12200547
references.citation: Appl Environ Microbiol. 2012 Apr;78(7):2337-44
references.pmid: 22247175
references.citation: Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2857-62
references.pmid: 16481624
references.citation: Microbiology (Reading). 2008 Oct;154(Pt 10):3212-3223
references.pmid: 18832326
references.citation: Microbiology (Reading). 2007 Apr;153(Pt 4):1042-1058
references.pmid: 17379713
references.citation: J Biol Chem. 2009 May 8;284(19):12989-97
references.pmid: 19269972
references.citation: Proc Natl Acad Sci U S A. 2009 Oct 6;106(40):17193-8
references.pmid: 19805152
references.citation: Infect Immun. 2010 Mar;78(3):1294-303
references.pmid: 20028807
references.citation: BMC Microbiol. 2012 Jul 07;12:135
references.pmid: 22769602
references.citation: Nucleic Acids Res. 2003 Nov 15;31(22):6516-23
references.pmid: 14602910
references.citation: Microb Cell Fact. 2011 Aug 30;10 Suppl 1:S2
references.pmid: 21995776
references.citation: Nat Rev Microbiol. 2011 Mar;9(3):166-76
references.pmid: 21326273
references.citation: Mol Microbiol. 2006 Jul;61(1):126-41
references.pmid: 16824100
references.citation: Appl Environ Microbiol. 2007 Jul;73(13):4359-64
references.pmid: 17483264
references.citation: J Bacteriol. 2005 Jul;187(13):4646-55
references.pmid: 15968076
references.citation: J Clin Invest. 2006 Oct;116(10):2799-807
references.pmid: 17016560
references.citation: PLoS One. 2012;7(11):e49048
references.pmid: 23145064
references.citation: Nucleic Acids Res. 2009 Jul;37(Web Server issue):W202-8
references.pmid: 19458158
references.citation: Trends Microbiol. 2004 May;12(5):228-34
references.pmid: 15120142