Fis Is Essential for Yersinia pseudotuberculosis Virulence and Protects against Reactive Oxygen Species Produced by Phagocytic Cells during Infection.
All three pathogenic Yersinia species share a conserved virulence plasmid that encodes a Type 3 Secretion System (T3SS) and its associated effector proteins. During mammalian infection, these effectors are injected into innate immune cells, where they block many bactericidal functions, including the production of reactive oxygen species (ROS). However, Y. pseudotuberculosis (Yptb) lacking the T3SS retains the ability to colonize host organs, demonstrating that chromosome-encoded factors are sufficient for growth within mammalian tissue sites. Previously we uncovered more than 30 chromosomal factors that contribute to growth of T3SS-deficient Yptb in livers. Here, a deep sequencing-based approach was used to validate and characterize the phenotype of 18 of these genes during infection by both WT and plasmid-deficient Yptb. Additionally, the fitness of these mutants was evaluated in immunocompromised mice to determine whether any genes contributed to defense against phagocytic cell restriction. Mutants containing deletions of the dusB-fis operon, which encodes the nucleoid associated protein Fis, were markedly attenuated in immunocompetent mice, but were restored for growth in mice lacking neutrophils and inflammatory monocytes, two of the major cell types responsible for restricting Yersinia infection. We determined that Fis was dispensable for secretion of T3SS effectors, but was essential for resisting ROS and regulated the transcription of several ROS-responsive genes. Strikingly, this protection was critical for virulence, as growth of ΔdusB-fis was restored in mice unable to produce ROS. These data support a model in which ROS generated by neutrophils and inflammatory monocytes that have not been translocated with T3SS effectors enter bacterial cells during infection, where their bactericidal effects are resisted in a Fis-dependent manner. This is the first report of the requirement for Fis during Yersinia infection and also highlights a novel mechanism by which Yptb defends against ROS in mammalian tissues.
Graduate Program in Molecular Microbiology, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts, United States of America.
Erin R
ER
Green
Graduate Program in Molecular Microbiology, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts, United States of America.
Stacie
S
Clark
Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America.
Gregory T
GT
Crimmins
Universitatsklinikum Regensburg, Innere Medizin II/Nephrologie-Transplantation, Regensburg, Germany.
Matthias
M
Mack
Graduate Program in Molecular Microbiology, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts, United States of America.|Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America.
Carol A
CA
Kumamoto
Graduate Program in Molecular Microbiology, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts, United States of America.|Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America.
Joan
J
Mecsas
Kidney Int. 2013 Jul;84(1):78-89
23486523
Infect Immun. 2008 May;76(5):2070-9
18285492
Am J Physiol Lung Cell Mol Physiol. 2005 Feb;288(2):L350-8
15516494
Immunol Cell Biol. 2015 Oct;93(9):815-24
25829141
Infect Immun. 2004 Feb;72(2):908-15
14742535
Proc Natl Acad Sci U S A. 2006 Sep 5;103(36):13514-9
16938880
Mol Microbiol. 2001 Oct;42(2):439-52
11703666
Infect Immun. 1995 Feb;63(2):681-5
7822039
Int Rev Cytol. 2005;246:135-88
16164968
Cell Host Microbe. 2013 Sep 11;14(3):306-17
24034616
J Bacteriol. 2012 May;194(10):2458-69
22389479
Nat Rev Microbiol. 2013 Jul;11(7):435-42
23712350
PLoS Pathog. 2006 Sep;2(9):e86
16948531
Arthritis Rheum. 2007 Sep;56(9):2975-85
17763443
PLoS Pathog. 2006 May;2(5):e42
16733541
Cell Mol Immunol. 2015 Jan;12(1):5-23
25263488
J Bacteriol. 1985 May;162(2):661-7
3886630
Infect Immun. 2007 Jan;75(1):429-42
17074849
Mol Microbiol. 2007 Feb;63(3):859-71
17181781
PLoS Pathog. 2015 Mar 12;11(3):e1004651
25764063
Adv Exp Med Biol. 2003;529:31-3
12756723
Proc Natl Acad Sci U S A. 2006 Aug 1;103(31):11766-71
16864791
Mol Microbiol. 1999 Apr;32(1):51-62
10216859
Curr Opin Microbiol. 2009 Feb;12(1):94-100
19185531
Cell Microbiol. 2014 Jul;16(7):1014-23
24779390
J Bacteriol. 1996 Sep;178(18):5347-52
8808921
Cell. 1985 Jul;41(3):753-62
2988786
Trends Microbiol. 2014 Feb;22(2):92-9
24370464
PLoS Pathog. 2013;9(6):e1003415
23818844
J Bacteriol. 1987 Jan;169(1):26-32
3539918
Infect Immun. 1999 Aug;67(8):3703-13
10417127
Mol Microbiol. 2000 Mar;35(6):1413-20
10760142
Infect Immun. 2010 Jun;78(6):2723-33
20385761
Infect Immun. 2001 Dec;69(12 ):7810-9
11705963
J Bacteriol. 1997 Sep;179(18):5967-70
9294462
Nat Rev Microbiol. 2016 Apr;14(4):221-34
26949049
Curr Opin Microbiol. 2016 Feb;29:56-62
26638030
Cell Host Microbe. 2016 Aug 10;20(2):133-43
27512901
J Bacteriol. 2001 Dec;183(24):7173-81
11717276
Infect Immun. 2001 May;69(5):2779-87
11292689
Environ Microbiol. 2009 Apr;11(4):992-1006
19187284
J Leukoc Biol. 2013 Sep;94(3):409-21
23825388
Microbiology. 2004 Jul;150(Pt 7):2037-53
15256548
Nat Immunol. 2007 Jan;8(1):3-4
17179960
Cell Microbiol. 2010 Aug;12(8):1064-82
20148898
J Bacteriol. 1989 Apr;171(4):2049-55
2649484
Nature. 1981 May 21;291(5812):238-9
7015147
J Immunol. 2001 Apr 1;166(7):4697-704
11254730
Cell Microbiol. 2010 Jul;12(7):988-1001
20148901
Microbiology. 2015 Jun;161(6):1282-94
25767108
J Immunol. 2008 Aug 15;181(4):2713-22
18684962
Cell Microbiol. 2014 Feb;16(2):247-68
24119087
PLoS Pathog. 2015 May 14;11(5):e1004893
25974210
Annu Rev Immunol. 2008;26:421-52
18303997
Infect Immun. 2004 Sep;72(9):4973-84
15321989
Cell Host Microbe. 2015 Jan 14;17(1):21-31
25500192
PLoS Pathog. 2012;8(8):e1002828
22876175
J Bacteriol. 1992 Dec;174(24):8043-56
1459953
Microbiol Rev. 1991 Dec;55(4):561-85
1779927
J Biochem Biophys Methods. 2006 Apr 30;67(1):67-74
16480772
J Med Microbiol. 1984 Dec;18(3):371-5
6389874
MBio. 2014 Jul 01;5(4):e01402-14
24987096
Mol Microbiol. 2011 Jul;81(1):179-91
21542864
Cell Host Microbe. 2012 Dec 13;12(6):799-805
23245324
Clin Microbiol Rev. 2007 Oct;20(4):535-49
17934073
Mol Microbiol. 2001 Jan;39(1):79-88
11123690
Biotechniques. 2013 Jan;54(1):25-34
23311318
MBio. 2014 Aug 19;5(4):null
25139902
Nat Genet. 1995 Feb;9(2):202-9
7719350
PLoS Pathog. 2010 Feb 05;6(2):e1000750
20140235
Tuber Lung Dis. 1997;78(5-6):237-46
10209678
J Biol Chem. 2003 May 23;278(21):19463-72
12637497
J Exp Med. 2000 Jul 17;192(2):237-48
10899910
Infect Immun. 1997 Nov;65(11):4705-10
9353054
J Biol Chem. 1986 Aug 15;261(23):10632-6
3015935
J Bacteriol. 1998 Nov;180(22):5932-46
9811652
Antimicrob Agents Chemother. 2010 Aug;54(8):3241-54
20498321
PLoS One. 2013 May 23;8(5):e64688
23717649
Infect Immun. 2003 Sep;71(9):5432-5
12933899
Antioxid Redox Signal. 2006 Sep-Oct;8(9-10):1573-82
16987012
Cell Microbiol. 2013 Oct;15(10):1622-31
23834311