title: Listeria monocytogenes PrsA2 is required for virulence factor secretion and bacterial viability within the host cell cytosol.
abstract: In the course of establishing its replication niche within the cytosol of infected host cells, the facultative intracellular bacterial pathogen Listeria monocytogenes must efficiently regulate the secretion and activity of multiple virulence factors. L. monocytogenes encodes two predicted posttranslocation secretion chaperones, PrsA1 and PrsA2, and evidence suggests that PrsA2 has been specifically adapted for bacterial pathogenesis. PrsA-like chaperones have been identified in a number of Gram-positive bacteria, where they are reported to function at the bacterial membrane-cell wall interface to assist in the folding of proteins translocated across the membrane; in some cases, these proteins have been found to be essential for bacterial viability. In this study, the contributions of PrsA2 and PrsA1 to L. monocytogenes growth and protein secretion were investigated in vitro and in vivo. Neither PrsA2 nor PrsA1 was found to be essential for L. monocytogenes growth in broth culture; however, optimal bacterial viability was found to be dependent upon PrsA2 for L. monocytogenes located within the cytosol of host cells. Proteomic analyses of prsA2 mutant strains in the presence of a mutationally activated allele of the virulence regulator PrfA revealed a critical requirement for PrsA2 activity under conditions of PrfA activation, an event which normally takes place within the host cell cytosol. Despite a high degree of amino acid similarity, no detectable degree of functional overlap was observed between PrsA2 and PrsA1. Our results indicate a critical requirement for PrsA2 under conditions relevant to host cell infection.
authors.affiliation: Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago, IL 60612-7344, USA.
authors.forename: Francis
authors.initials: F
authors.lastname: Alonzo
authors.forename: Nancy E
authors.initials: NE
authors.lastname: Freitag
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D002460:Cell Line; D003600:Cytosol / Q000382:microbiology*; D005260:Female; D005347:Fibroblasts / Q000382:microbiology; D015964:Gene Expression Regulation, Bacterial; D008089:Listeria monocytogenes / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D008088:Listeriosis / Q000382:microbiology*; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D050296:Microbial Viability; D019696:Peptidylprolyl Isomerase / Q000235:genetics / Q000378:metabolism*; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D037521:Virulence Factors; D019696:Peptidylprolyl Isomerase; C541924:PrsA2 protein, Listeria monocytogenes
references.citation: Infect Immun. 2007 Nov;75(11):5135-47
references.pmid: 17682039
references.citation: Infect Immun. 1993 Apr;61(4):1576-80
references.pmid: 8384163
references.citation: J Cell Biol. 1997 Jun 16;137(6):1381-92
references.pmid: 9182669
references.citation: Nat Rev Microbiol. 2008 Apr;6(4):276-87
references.pmid: 18327271
references.citation: Proc Natl Acad Sci U S A. 1999 May 25;96(11):6456-61
references.pmid: 10339609
references.citation: J Biol Chem. 2006 Oct 20;281(42):31812-22
references.pmid: 16905535
references.citation: Infect Immun. 2001 Aug;69(8):4938-43
references.pmid: 11447171
references.citation: Infect Immun. 1990 Nov;58(11):3770-8
references.pmid: 2172168
references.citation: Mol Microbiol. 2003 Jun;48(6):1537-51
references.pmid: 12791137
references.citation: Infect Immun. 2007 Jan;75(1):44-51
references.pmid: 17060464
references.citation: Appl Environ Microbiol. 2004 Jun;70(6):3457-66
references.pmid: 15184144
references.citation: Antimicrob Agents Chemother. 1990 Apr;34(4):539-42
references.pmid: 2111657
references.citation: Infect Immun. 1995 Oct;63(10):3896-903
references.pmid: 7558297
references.citation: Mol Microbiol. 2010 Jul 1;77(1):108-27
references.pmid: 20487272
references.citation: Mol Microbiol. 2009 Nov;74(4):956-73
references.pmid: 19818015
references.citation: Infect Immun. 2007 Dec;75(12):5886-97
references.pmid: 17938228
references.citation: Appl Environ Microbiol. 2006 Oct;72(10):6623-31
references.pmid: 17021213
references.citation: Proteomics. 2009 May;9(9):2432-43
references.pmid: 19343722
references.citation: Cell Microbiol. 2004 Mar;6(3):235-42
references.pmid: 14764107
references.citation: Infect Immun. 1970 Apr;1(4):363-72
references.pmid: 16557743
references.citation: Infect Immun. 2003 Dec;71(12):6754-65
references.pmid: 14638761
references.citation: J Bacteriol. 2009 Jun;191(12):3950-64
references.pmid: 19376879
references.citation: Infect Immun. 2006 Jan;74(1):765-8
references.pmid: 16369036
references.citation: Proteomics. 2005 Apr;5(6):1544-57
references.pmid: 15838904
references.citation: Cell. 1991 Jun 28;65(7):1127-41
references.pmid: 1905979
references.citation: J Bacteriol. 2003 Nov;185(21):6295-307
references.pmid: 14563864
references.citation: Infect Immun. 2006 May;74(5):2505-12
references.pmid: 16622185
references.citation: Infect Immun. 1997 Jan;65(1):101-9
references.pmid: 8975898
references.citation: Mol Microbiol. 2008 Dec;70(6):1342-57
references.pmid: 19019159
references.citation: Nat Rev Microbiol. 2009 Sep;7(9):623-8
references.pmid: 19648949
references.citation: J Biol Chem. 1975 May 25;250(10):4007-21
references.pmid: 236308
references.citation: J Bacteriol. 1997 Mar;179(5):1533-40
references.pmid: 9045810
references.citation: Proteomics. 2009 Jun;9(11):3136-55
references.pmid: 19526548
references.citation: J Cell Biol. 2000 Aug 7;150(3):527-38
references.pmid: 10931865
references.citation: Mol Microbiol. 2000 Jan;35(2):289-98
references.pmid: 10652090
references.citation: APMIS. 1989 Nov;97(11):1013-7
references.pmid: 2511904
references.citation: Infect Immun. 2006 Feb;74(2):1323-38
references.pmid: 16428782
references.citation: Infect Immun. 2009 Jul;77(7):2612-23
references.pmid: 19451247
references.citation: Infect Immun. 2004 Dec;72(12):7374-8
references.pmid: 15557671
references.citation: Appl Environ Microbiol. 2000 Apr;66(4):1405-9
references.pmid: 10742219
references.citation: Infect Immun. 2001 Sep;69(9):5953-7
references.pmid: 11500481
references.citation: Proteomics. 2004 Oct;4(10):3187-201
references.pmid: 15378702
references.citation: Mol Microbiol. 2006 Dec;62(6):1631-42
references.pmid: 17427287
references.citation: Genes Dev. 2007 Oct 1;21(19):2473-84
references.pmid: 17908933
references.citation: Microbes Infect. 2007 Aug;9(10):1176-87
references.pmid: 17720603
references.citation: Clin Microbiol Rev. 2001 Jul;14(3):584-640
references.pmid: 11432815
references.citation: Infect Immun. 1989 Aug;57(8):2569-73
references.pmid: 2545629
references.citation: Microbiology (Reading). 2003 Mar;149(Pt 3):569-577
references.pmid: 12634326
references.citation: FEMS Microbiol Lett. 2005 Oct 15;251(2):281-8
references.pmid: 16140473
references.citation: Microbes Infect. 2007 Aug;9(10):1156-66
references.pmid: 17764999
references.citation: Acta Microbiol Hung. 1990;37(2):227-31
references.pmid: 2125398
references.citation: FEMS Microbiol Lett. 1991 Nov 15;68(2):191-5
references.pmid: 1778442
references.citation: Mol Microbiol. 1994 Jun;12(5):845-53
references.pmid: 8052135
references.citation: Microb Drug Resist. 2001 Summer;7(2):127-35
references.pmid: 11442339
references.citation: J Pathol. 2006 Jan;208(2):215-23
references.pmid: 16362984
references.citation: Proteomics. 2004 Oct;4(10):2991-3006
references.pmid: 15378750
references.citation: Future Microbiol. 2006 Jun;1(1):89-101
references.pmid: 17661688
references.citation: Appl Environ Microbiol. 2004 Mar;70(3):1669-79
references.pmid: 15006792
references.citation: EMBO Rep. 2008 Aug;9(8):735-41
references.pmid: 18617888
references.citation: Mol Cell. 2002 Sep;10(3):443-55
references.pmid: 12408815
references.citation: Mol Microbiol. 1993 Sep;9(5):1097-106
references.pmid: 7934915
references.citation: J Bacteriol. 2001 Mar;183(6):1881-90
references.pmid: 11222585
references.citation: J Bacteriol. 2004 Sep;186(18):6265-76
references.pmid: 15342597
references.citation: FEMS Microbiol Rev. 2006 Sep;30(5):774-805
references.pmid: 16911044
references.citation: Infect Immun. 1992 Mar;60(3):916-21
references.pmid: 1311708
references.citation: Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11890-4
references.pmid: 8265643
references.citation: Infect Immun. 1995 Nov;63(11):4231-7
references.pmid: 7591052
references.citation: Mol Microbiol. 2000 Mar;35(6):1286-94
references.pmid: 10760131
references.citation: Cell Microbiol. 2007 Jun;9(6):1365-75
references.pmid: 17419717
references.citation: Gene. 2008 Feb 29;410(1):113-21
references.pmid: 18222616
references.citation: Trends Microbiol. 2009 Apr;17(4):139-45
references.pmid: 19299134
references.citation: J Bacteriol. 2005 Apr;187(8):2601-8
references.pmid: 15805506
references.citation: Appl Environ Microbiol. 2008 Jun;74(12):3823-30
references.pmid: 18424542
references.citation: Cell Microbiol. 2004 Feb;6(2):155-66
references.pmid: 14706101
references.citation: Int J Med Microbiol. 2007 Oct;297(6):401-15
references.pmid: 17482513
references.citation: Appl Environ Microbiol. 2002 Apr;68(4):1697-705
references.pmid: 11916687
references.citation: Antimicrob Agents Chemother. 2006 Aug;50(8):2824-8
references.pmid: 16870778
references.citation: Microbes Infect. 2007 Aug;9(10):1167-75
references.pmid: 17761447
references.citation: Microbes Infect. 2007 Aug;9(10):1196-207
references.pmid: 17764998
references.citation: Mol Microbiol. 1996 Aug;21(4):823-37
references.pmid: 8878044
references.citation: Biochemistry. 2001 Sep 25;40(38):11390-404
references.pmid: 11560487
references.citation: Gut Pathog. 2009 Dec 15;1:23
references.pmid: 20003484
references.citation: Infect Immun. 1995 Nov;63(11):4531-4
references.pmid: 7591098
references.citation: Proc Natl Acad Sci U S A. 1989 Jul;86(14):5522-6
references.pmid: 2501788
references.citation: EMBO J. 1992 May;11(5):1981-90
references.pmid: 1582425
references.citation: Antimicrob Agents Chemother. 1990 Sep;34(9):1695-8
references.pmid: 2126687
references.citation: J Biol Chem. 2004 Apr 30;279(18):19302-14
references.pmid: 14976191
references.citation: Infect Immun. 2003 Jun;71(6):3614-8
references.pmid: 12761148
references.citation: Microbiology (Reading). 2008 Nov;154(Pt 11):3579-3589
references.pmid: 18957610
references.citation: Nature. 2000 Aug 17;406(6797):768-74
references.pmid: 10963606
references.citation: J Bacteriol. 1980 Feb;141(2):876-87
references.pmid: 6767692
references.citation: Mol Microbiol. 2007 Mar;63(5):1453-67
references.pmid: 17302820
references.citation: Trends Microbiol. 1997 Jul;5(7):272-6
references.pmid: 9234509
references.citation: Mol Microbiol. 2009 Oct;74(2):299-314
references.pmid: 19682249
references.citation: Biochim Biophys Acta. 2004 Nov 11;1694(1-3):311-27
references.pmid: 15546674
references.citation: Appl Environ Microbiol. 1998 Sep;64(9):3147-52
references.pmid: 9726852
references.citation: J Antimicrob Chemother. 1990 May;25(5):751-8
references.pmid: 2115510
references.citation: Cell. 1992 Feb 7;68(3):521-31
references.pmid: 1739966
references.citation: Infect Immun. 1992 Jan;60(1):219-30
references.pmid: 1309513
references.citation: Mol Microbiol. 2001 Sep;41(5):1159-72
references.pmid: 11555295
references.citation: J Mol Biol. 1998 Jun 12;279(3):485-99
references.pmid: 9641973
references.citation: J Bacteriol. 2001 Dec;183(23):6794-800
references.pmid: 11698367
references.citation: Mol Microbiol. 2006 Aug;61(4):999-1012
references.pmid: 16859495
references.citation: Appl Environ Microbiol. 2005 Aug;71(8):4241-7
references.pmid: 16085809
references.citation: Microb Pathog. 2008 Oct;45(4):273-81
references.pmid: 18675335
references.citation: J Cell Biol. 2002 Mar 18;156(6):1029-38
references.pmid: 11901168
references.citation: Acta Microbiol Pol. 2003;52(2):131-42
references.pmid: 14594400
references.citation: J Bacteriol. 2002 Aug;184(15):4177-86
references.pmid: 12107135
references.citation: Cell Microbiol. 2006 Jan;8(1):107-19
references.pmid: 16367870
references.citation: Nature. 1992 May 21;357(6375):257-60
references.pmid: 1589024
references.citation: Appl Environ Microbiol. 1994 Jul;60(7):2360-6
references.pmid: 8074517
references.citation: Microbiology (Reading). 2005 Oct;151(Pt 10):3215-3222
references.pmid: 16207905
references.citation: J Cell Biol. 2001 Oct 1;155(1):89-100
references.pmid: 11581288
references.citation: Infect Immun. 2005 Apr;73(4):1917-26
references.pmid: 15784531
references.citation: Arch Microbiol. 2000 Jan;173(1):21-8
references.pmid: 10648100
references.citation: Acta Microbiol Pol. 2002;51(1):5-12
references.pmid: 12184448
references.citation: Appl Environ Microbiol. 2004 Apr;70(4):1935-43
references.pmid: 15066783
references.citation: Curr Opin Microbiol. 2010 Feb;13(1):47-52
references.pmid: 20015680
references.citation: Infect Immun. 2000 Dec;68(12):7069-77
references.pmid: 11083832
references.citation: Appl Environ Microbiol. 2002 Apr;68(4):1491-8
references.pmid: 11916660