title: Role of lipid A acylation in Yersinia enterocolitica virulence.
abstract: Yersinia enterocolitica is an important human pathogen. Y. enterocolitica must adapt to the host environment, and temperature is an important cue regulating the expression of most Yersinia virulence factors. Here, we report that Y. enterocolitica 8081 serotype O:8 synthesized tetra-acylated lipid A at 37 degrees C but that hexa-acylated lipid A predominated at 21 degrees C. By mass spectrometry and genetic methods, we have shown that the Y. enterocolitica msbB, htrB, and lpxP homologues encode the acyltransferases responsible for the addition of C(12), C(14) and C(16:1), respectively, to lipid A. The expression levels of the acyltransferases were temperature regulated. Levels of expression of msbB and lpxP were higher at 21 degrees C than at 37 degrees C, whereas the level of expression of htrB was higher at 37 degrees C. At 21 degrees C, an lpxP mutant was the strain most susceptible to polymyxin B, whereas at 37 degrees C, an htrB mutant was the most susceptible. We present evidence that the lipid A acylation status affects the expression of Yersinia virulence factors. Thus, expression of flhDC, the flagellar master regulatory operon, was downregulated in msbB and lpxP mutants, with a concomitant decrease in motility. Expression of the phospholipase yplA was also downregulated in both mutants. inv expression was downregulated in msbB and htrB mutants, and consistent with this finding, invasion of HeLa cells was diminished. However, the expression of rovA, the positive regulator of inv, was not affected in the mutants. The levels of pYV-encoded virulence factors Yops and YadA in the acyltransferase mutants were not affected. Finally, we show that only the htrB mutant was attenuated in vivo.
authors.affiliation: Fundación Caubet- CIMERA Illes Balears, 07110 Bunyola, Spain.
authors.forename: Camino
authors.initials: C
authors.lastname: Pérez-Gutiérrez
authors.forename: Enrique
authors.initials: E
authors.lastname: Llobet
authors.forename: Catalina M
authors.initials: CM
authors.lastname: Llompart
authors.forename: Mar
authors.initials: M
authors.lastname: Reinés
authors.forename: José A
authors.initials: JA
authors.lastname: Bengoechea
mesh_terms: D000215:Acylation; D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism; D005408:Flagellin / Q000096:biosynthesis; D017353:Gene Deletion; D020869:Gene Expression Profiling; D006367:HeLa Cells; D006801:Humans; D008050:Lipid A / Q000737:chemistry / Q000378:metabolism*; D008124:Locomotion; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D011112:Polymyxin B / Q000494:pharmacology; D013696:Temperature; D014774:Virulence; D037521:Virulence Factors / Q000378:metabolism*; D015009:Yersinia Infections / Q000382:microbiology / Q000473:pathology; D015008:Yersinia enterocolitica / Q000187:drug effects / Q000472:pathogenicity* / Q000528:radiation effects
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D008050:Lipid A; D037521:Virulence Factors; D005408:Flagellin; D000217:Acyltransferases; D011112:Polymyxin B
references.citation: J Biol Chem. 2005 Aug 5;280(31):28186-94
references.pmid: 15951433
references.citation: J Bacteriol. 1996 Dec;178(23):6857-64
references.pmid: 8955307
references.citation: J Bacteriol. 2006 Feb;188(4):1381-8
references.pmid: 16452420
references.citation: Mol Microbiol. 1998 Mar;27(6):1171-82
references.pmid: 9570402
references.citation: J Bacteriol. 2006 Jul;188(14):5101-12
references.pmid: 16816182
references.citation: J Endotoxin Res. 2005;11(3):167-73
references.pmid: 15949145
references.citation: Microbiology (Reading). 2007 Aug;153(Pt 8):2416-2425
references.pmid: 17660406
references.citation: Nat Rev Mol Cell Biol. 2002 Oct;3(10):742-52
references.pmid: 12360191
references.citation: Methods Mol Biol. 2000;145:311-25
references.pmid: 10820729
references.citation: Int J Med Microbiol. 2001 Aug;291(3):209-18
references.pmid: 11554561
references.citation: J Biol Chem. 2002 Apr 19;277(16):14186-93
references.pmid: 11830594
references.citation: Proc Natl Acad Sci U S A. 1999 May 25;96(11):6456-61
references.pmid: 10339609
references.citation: Mol Microbiol. 1994 Jan;11(1):123-35
references.pmid: 7511772
references.citation: Infect Immun. 2007 Mar;75(3):1512-6
references.pmid: 17178779
references.citation: Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6473-7
references.pmid: 8341658
references.citation: Mol Microbiol. 1998 Jul;29(2):571-9
references.pmid: 9720873
references.citation: Cell. 1998 Oct 16;95(2):189-98
references.pmid: 9790526
references.citation: J Biol Chem. 1998 Dec 4;273(49):32369-72
references.pmid: 9829962
references.citation: Mol Microbiol. 2004 Apr;52(2):451-69
references.pmid: 15066033
references.citation: Int J Med Microbiol. 2002 Feb;291(6-7):455-62
references.pmid: 11890544
references.citation: J Infect Dis. 2007 Oct 1;196(7):1088-92
references.pmid: 17763333
references.citation: Infect Immun. 1997 Nov;65(11):4778-83
references.pmid: 9353064
references.citation: Infect Immun. 1990 Sep;58(9):2840-9
references.pmid: 2129533
references.citation: Antimicrob Agents Chemother. 2003 Oct;47(10):3332-5
references.pmid: 14506051
references.citation: J Bacteriol. 1999 May;181(9):2823-33
references.pmid: 10217774
references.citation: Mol Microbiol. 2002 May;44(4):1045-62
references.pmid: 12010497
references.citation: Mol Microbiol. 2004 Jun;52(5):1363-73
references.pmid: 15165239
references.citation: Infect Immun. 2000 Jun;68(6):3768-71
references.pmid: 10816545
references.citation: Science. 1994 Apr 15;264(5157):382-8
references.pmid: 8153625
references.citation: Trends Microbiol. 1994 Nov;2(11):444-9
references.pmid: 7866702
references.citation: J Biol Chem. 2006 Aug 4;281(31):21974-21987
references.pmid: 16704973
references.citation: Appl Environ Microbiol. 2008 Feb;74(3):633-44
references.pmid: 18065606
references.citation: Curr Issues Mol Biol. 2005 Jul;7(2):151-77
references.pmid: 16053248
references.citation: Curr Issues Mol Biol. 2006 Jan;8(1):11-26
references.pmid: 16450883
references.citation: J Endotoxin Res. 2003;9(2):67-84
references.pmid: 12803879
references.citation: Adv Exp Med Biol. 2003;529:267-74
references.pmid: 12756770
references.citation: Infect Immun. 2007 Sep;75(9):4386-99
references.pmid: 17620356
references.citation: J Biol Chem. 2007 May 25;282(21):15569-77
references.pmid: 17371870
references.citation: Infect Immun. 1988 May;56(5):1242-8
references.pmid: 2833444
references.citation: J Bacteriol. 2005 Nov;187(21):7407-16
references.pmid: 16237024
references.citation: Infect Immun. 1999 Dec;67(12):6583-90
references.pmid: 10569778
references.citation: FEBS Lett. 2000 Jan 7;465(1):87-92
references.pmid: 10620712
references.citation: Genetics. 1954 Jul;39(4):440-52
references.pmid: 17247495
references.citation: Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129-33
references.pmid: 2999794
references.citation: Trends Microbiol. 1995 Aug;3(8):310-7
references.pmid: 8528615
references.citation: Infect Immun. 2002 Aug;70(8):4165-76
references.pmid: 12117925
references.citation: Microbiol Mol Biol Rev. 2003 Dec;67(4):593-656
references.pmid: 14665678
references.citation: J Bacteriol. 2004 Jul;186(13):4124-33
references.pmid: 15205413
references.citation: Science. 2005 Feb 25;307(5713):1313-7
references.pmid: 15731456
references.citation: J Bacteriol. 2007 Mar;189(6):2203-9
references.pmid: 17209027
references.citation: Genome Res. 1995 Nov;5(4):404-7
references.pmid: 8750200
references.citation: J Bacteriol. 2008 Nov;190(21):7012-21
references.pmid: 18757539
references.citation: Clin Microbiol Rev. 1997 Apr;10(2):257-76
references.pmid: 9105754
references.citation: PLoS Genet. 2006 Dec 15;2(12):e206
references.pmid: 17173484
references.citation: Annu Rev Biochem. 2002;71:635-700
references.pmid: 12045108
references.citation: J Bacteriol. 2002 Mar;184(5):1324-34
references.pmid: 11844761
references.citation: Infect Immun. 1997 Nov;65(11):4431-5
references.pmid: 9353016
references.citation: J Biol Chem. 1980 Apr 25;255(8):3263-5
references.pmid: 6988423
references.citation: Mol Microbiol. 1995 Aug;17(3):575-94
references.pmid: 8559076
references.citation: J Immunol Methods. 1991 Mar 21;137(2):167-73
references.pmid: 1901580
references.citation: Trends Microbiol. 2002 Apr;10(4):179-86
references.pmid: 11912025
references.citation: Mol Microbiol. 2005 Mar;55(5):1403-12
references.pmid: 15720549
references.citation: Infect Immun. 1998 Aug;66(8):3941-51
references.pmid: 9673284
references.citation: Mol Microbiol. 2000 Feb;35(3):677-85
references.pmid: 10672189
references.citation: Infect Immun. 2002 Aug;70(8):4092-8
references.pmid: 12117916
references.citation: J Bacteriol. 1998 Feb;180(4):793-800
references.pmid: 9473031
references.citation: Appl Environ Microbiol. 1998 Jul;64(7):2710-5
references.pmid: 9647854
references.citation: J Bacteriol. 2000 Apr;182(8):2314-20
references.pmid: 10735878
references.citation: Science. 1999 Nov 19;286(5444):1561-5
references.pmid: 10567263
references.citation: Science. 1997 Apr 11;276(5310):250-3
references.pmid: 9092473
references.citation: Infect Immun. 2003 Apr;71(4):2014-21
references.pmid: 12654821
references.citation: J Immunol. 2002 May 15;168(10):5240-51
references.pmid: 11994481
references.citation: J Bacteriol. 1992 Mar;174(6):2047-51
references.pmid: 1548243
references.citation: Biotechniques. 1999 May;26(5):824-6, 828
references.pmid: 10337469
references.citation: Mol Microbiol. 1997 Jan;23(1):63-76
references.pmid: 9004221
references.citation: EMBO J. 2004 Aug 4;23(15):2931-41
references.pmid: 15272304
references.citation: Eur J Biochem. 2001 Feb;268(3):554-64
references.pmid: 11168394
references.citation: J Lipid Res. 2005 Aug;46(8):1773-8
references.pmid: 15930524
references.citation: Gene. 1991 Dec 20;109(1):137-41
references.pmid: 1756974