Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris.
Lipoprotein
Proteomic analysis
Virulence
Journal
BMC microbiology
ISSN: 1471-2180
Titre abrégé: BMC Microbiol
Pays: England
ID NLM: 100966981
Informations de publication
Date de publication:
21 01 2019
21 01 2019
Historique:
received:
13
09
2018
accepted:
02
01
2019
entrez:
23
1
2019
pubmed:
23
1
2019
medline:
24
9
2019
Statut:
epublish
Résumé
The gram-negative Xanthomonas campestris pv. campestris is the pathogenic bacterium that causes black rot disease in crucifers. The virulence determinants of this bacterium include extracellular enzymes, exopolysaccharides, and biofilm formation. Here, one transposon mutant of X. campestris pv. campestris strain 17 that affects biofilm formation was isolated, and subsequent analyses led to the identification of the lolA gene, which encodes an outer membrane lipoprotein chaperone. The lolA mutant exhibited significant reductions in bacterial attachment, extracellular enzyme production, virulence, and tolerance in the presence of myriad membrane-perturbing agents. These phenotypic changes of the mutant could be complemented to the wild-type level through the intact lolA gene. Proteomic analysis revealed that 109 proteins were differentially expressed after lolA mutation. These differentially expressed proteins were categorized in various functional groups and were mainly associated with the membrane component, were involved in transport, and contained receptor activity. Through reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis, deletion of lolA was determined to have caused significantly reduced expression of genes that encode the major extracellular enzymes, the biofilm-related proteins, and the virulence-related proteins. The RT-qPCR analysis also indicated that the expression of several genes that encode putative outer membrane lipoproteins and TonB-dependent receptors was reduced after lolA mutation. This is the first report to define the lolA gene as a virulence factor and to contribute to the functional understanding of, and provide new information concerning, the role of lolA in Xanthomonas. Furthermore, the results of this study provide and extend new insights into the function of lolA in bacteria.
Sections du résumé
BACKGROUND
The gram-negative Xanthomonas campestris pv. campestris is the pathogenic bacterium that causes black rot disease in crucifers. The virulence determinants of this bacterium include extracellular enzymes, exopolysaccharides, and biofilm formation. Here, one transposon mutant of X. campestris pv. campestris strain 17 that affects biofilm formation was isolated, and subsequent analyses led to the identification of the lolA gene, which encodes an outer membrane lipoprotein chaperone.
RESULTS
The lolA mutant exhibited significant reductions in bacterial attachment, extracellular enzyme production, virulence, and tolerance in the presence of myriad membrane-perturbing agents. These phenotypic changes of the mutant could be complemented to the wild-type level through the intact lolA gene. Proteomic analysis revealed that 109 proteins were differentially expressed after lolA mutation. These differentially expressed proteins were categorized in various functional groups and were mainly associated with the membrane component, were involved in transport, and contained receptor activity. Through reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis, deletion of lolA was determined to have caused significantly reduced expression of genes that encode the major extracellular enzymes, the biofilm-related proteins, and the virulence-related proteins. The RT-qPCR analysis also indicated that the expression of several genes that encode putative outer membrane lipoproteins and TonB-dependent receptors was reduced after lolA mutation.
CONCLUSIONS
This is the first report to define the lolA gene as a virulence factor and to contribute to the functional understanding of, and provide new information concerning, the role of lolA in Xanthomonas. Furthermore, the results of this study provide and extend new insights into the function of lolA in bacteria.
Identifiants
pubmed: 30665348
doi: 10.1186/s12866-019-1387-9
pii: 10.1186/s12866-019-1387-9
pmc: PMC6341742
doi:
Substances chimiques
Bacterial Proteins
0
Periplasmic Binding Proteins
0
Proteome
0
Virulence Factors
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
20Références
FEMS Microbiol Lett. 2014 Sep;358(1):81-90
pubmed: 25039264
Microbes Infect. 2004 May;6(6):623-9
pubmed: 15158198
Arch Microbiol. 2014 Oct;196(10):729-38
pubmed: 25033927
Nat Methods. 2011 Sep 29;8(10):785-6
pubmed: 21959131
BMC Bioinformatics. 2005 Jul 02;6:167
pubmed: 15992409
Biochim Biophys Acta. 2004 Jul 23;1693(1):5-13
pubmed: 15276320
Biotechnol Adv. 1999 Nov;17(6):489-508
pubmed: 14538126
Virulence. 2014;5(8):835-51
pubmed: 25603429
J Bacteriol. 2011 Oct;193(19):5450-64
pubmed: 21784931
EMBO J. 2013 Sep 11;32(18):2430-8
pubmed: 23881098
Proteomics. 2005 Jan;5(1):153-67
pubmed: 15619296
Res Microbiol. 2016 May;167(4):299-312
pubmed: 26804425
Genome Announc. 2017 Feb 23;5(8):
pubmed: 28232425
J Bacteriol. 2006 Apr;188(8):2761-73
pubmed: 16585737
Genome Announc. 2015 Dec 17;3(6):
pubmed: 26679582
Nat Protoc. 2009;4(1):44-57
pubmed: 19131956
PLoS One. 2007 Feb 21;2(2):e224
pubmed: 17311090
Sci Rep. 2015 Dec 01;5:17593
pubmed: 26621210
BMC Genomics. 2015 Nov 18;16:975
pubmed: 26581393
Trends Biochem Sci. 2017 Mar;42(3):232-242
pubmed: 27839654
Microbiology. 2013 Sep;159(Pt 9):1911-1919
pubmed: 23813675
J Biotechnol. 2013 Aug 20;167(2):111-22
pubmed: 23792782
Microbiology. 2010 Sep;156(Pt 9):2842-2854
pubmed: 20522496
Proteomics. 2007 Jun;7(12):2047-58
pubmed: 17566974
Arch Microbiol. 2004 Sep;182(1):1-6
pubmed: 15221203
FEMS Microbiol Rev. 2008 Aug;32(5):842-57
pubmed: 18557946
Mol Cell Proteomics. 2010 Dec;9(12):2704-18
pubmed: 20823122
BMC Microbiol. 2008 Jun 02;8:87
pubmed: 18518965
J Biotechnol. 2008 Mar 20;134(1-2):33-45
pubmed: 18304669
J Agric Food Chem. 2010 Feb 10;58(3):1653-63
pubmed: 20073482
Nucleic Acids Res. 2005 Jan 26;33(2):577-86
pubmed: 15673718
Biochem Biophys Res Commun. 2002 Jul 5;295(1):43-9
pubmed: 12083764
FEBS Lett. 1998 Nov 13;439(1-2):51-4
pubmed: 9849875
Biochim Biophys Acta. 2014 Aug;1843(8):1509-16
pubmed: 24780125
Nucleic Acids Res. 2014 Jan;42(Database issue):D222-30
pubmed: 24288371
Arch Microbiol. 2017 Aug;199(6):917-929
pubmed: 28378142
Nat Rev Microbiol. 2011 May;9(5):344-55
pubmed: 21478901
J Bacteriol. 2010 Nov;192(21):5657-62
pubmed: 20802033
J Mass Spectrom. 2001 Oct;36(10):1083-91
pubmed: 11747101
EMBO J. 1995 Jul 17;14(14):3365-72
pubmed: 7628437
Appl Environ Microbiol. 2006 Sep;72(9):6212-24
pubmed: 16957248
FEBS Lett. 2008 Jun 25;582(15):2247-51
pubmed: 18503771
J Bacteriol. 2015 May;197(10):1705-15
pubmed: 25755189
J Clin Microbiol. 1986 Sep;24(3):330-2
pubmed: 3489731
Microbiol Res. 2014 May-Jun;169(5-6):441-52
pubmed: 24120348
J Bacteriol. 2011 Nov;193(22):6342-57
pubmed: 21908674
Genome Res. 2005 Jun;15(6):757-67
pubmed: 15899963
Mol Microbiol. 2004 May;52(3):613-9
pubmed: 15101969
BMC Microbiol. 2005 Oct 07;5:58
pubmed: 16212653
J Basic Microbiol. 2018 May;58(5):403-413
pubmed: 29504631
Proc Natl Acad Sci U S A. 2017 May 2;114(18):4769-4774
pubmed: 28416660
J Bacteriol. 2012 Jul;194(14):3643-50
pubmed: 22563052
Nat Protoc. 2006;1(6):2856-60
pubmed: 17406544
Mol Plant Microbe Interact. 2008 Feb;21(2):151-61
pubmed: 18184059
Mol Plant Microbe Interact. 2012 Jan;25(1):69-84
pubmed: 21899385
J Biol Chem. 2005 Oct 14;280(41):34481-8
pubmed: 16091355
Mol Plant Pathol. 2013 Jan;14(1):2-18
pubmed: 23051837
BMC Microbiol. 2009 Jan 16;9:12
pubmed: 19149882
Lett Appl Microbiol. 1992 Feb;14(2):65-8
pubmed: 1367905
Philos Trans R Soc Lond B Biol Sci. 2015 Oct 5;370(1679):
pubmed: 26370942
Gene. 1988 Oct 15;70(1):191-7
pubmed: 2853689
J Biol Chem. 2007 May 4;282(18):13379-84
pubmed: 17350955
Nature. 2002 May 23;417(6887):459-63
pubmed: 12024217
J Bacteriol. 2011 Jan;193(1):246-64
pubmed: 20971899
FEMS Microbiol Rev. 2010 Mar;34(2):107-33
pubmed: 19925633
Biochim Biophys Acta Mol Cell Biol Lipids. 2017 Nov;1862(11):1414-1423
pubmed: 27871940
FEBS Lett. 2002 Sep 25;528(1-3):193-6
pubmed: 12297303
J Genet Genomics. 2013 Sep 20;40(9):473-87
pubmed: 24053949
J Mol Biol. 2010 Sep 3;401(5):921-30
pubmed: 20620146
J Agric Food Chem. 2009 Jul 22;57(14):6207-15
pubmed: 19601664
Proc Natl Acad Sci U S A. 2003 Sep 16;100(19):10995-1000
pubmed: 12960398
Methods Mol Biol. 2013;1007:183-217
pubmed: 23666727
Annu Rev Microbiol. 2011;65:239-59
pubmed: 21663440
Appl Environ Microbiol. 1990 Apr;56(4):919-23
pubmed: 2111116
FEBS Lett. 2005 Jul 4;579(17):3525-33
pubmed: 15955530
J Bacteriol. 2005 Nov;187(21):7254-66
pubmed: 16237009
Biochem Biophys Res Commun. 2001 Oct 12;287(5):1125-8
pubmed: 11587539
Microbiol Res. 2011 Oct 20;166(7):548-65
pubmed: 21237626