Dual Expression Profile of Type VI Secretion System Immunity Genes Protects Pandemic
The Vibrio cholerae type VI secretion system (T6SS) assembles as a molecular syringe that injects toxic protein effectors into both eukaryotic and prokaryotic cells. We previously reported that the V. cholerae O37 serogroup strain V52 maintains a constitutively active T6SS to kill other Gram-negative bacteria while being immune to attack by kin bacteria. The pandemic O1 El Tor V. cholerae strain C6706 is T6SS-silent under laboratory conditions as it does not produce T6SS structural components and effectors, and fails to kill Escherichia coli prey. Yet, C6706 exhibits full resistance when approached by T6SS-active V52. These findings suggested that an active T6SS is not required for immunity against T6SS-mediated virulence. Here, we describe a dual expression profile of the T6SS immunity protein-encoding genes tsiV1, tsiV2, and tsiV3 that provides pandemic V. cholerae strains with T6SS immunity and allows T6SS-silent strains to maintain immunity against attacks by T6SS-active bacterial neighbors. The dual expression profile allows transcription of the three genes encoding immunity proteins independently of other T6SS proteins encoded within the same operon. One of these immunity proteins, TsiV2, protects against the T6SS effector VasX which is encoded immediately upstream of tsiV2. VasX is a secreted, lipid-binding protein that we previously characterized with respect to T6SS-mediated virulence towards the social amoeba Dictyostelium discoideum. Our data suggest the presence of an internal promoter in the open reading frame of vasX that drives expression of the downstream gene tsiV2. Furthermore, VasX is shown to act in conjunction with VasW, an accessory protein to VasX, to compromise the inner membrane of prokaryotic target cells. The dual regulatory profile of the T6SS immunity protein-encoding genes tsiV1, tsiV2, and tsiV3 permits V. cholerae to tightly control T6SS gene expression while maintaining immunity to T6SS activity.
Vyšlo v časopise:
Dual Expression Profile of Type VI Secretion System Immunity Genes Protects Pandemic. PLoS Pathog 9(12): e32767. doi:10.1371/journal.ppat.1003752
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1003752
Souhrn
The Vibrio cholerae type VI secretion system (T6SS) assembles as a molecular syringe that injects toxic protein effectors into both eukaryotic and prokaryotic cells. We previously reported that the V. cholerae O37 serogroup strain V52 maintains a constitutively active T6SS to kill other Gram-negative bacteria while being immune to attack by kin bacteria. The pandemic O1 El Tor V. cholerae strain C6706 is T6SS-silent under laboratory conditions as it does not produce T6SS structural components and effectors, and fails to kill Escherichia coli prey. Yet, C6706 exhibits full resistance when approached by T6SS-active V52. These findings suggested that an active T6SS is not required for immunity against T6SS-mediated virulence. Here, we describe a dual expression profile of the T6SS immunity protein-encoding genes tsiV1, tsiV2, and tsiV3 that provides pandemic V. cholerae strains with T6SS immunity and allows T6SS-silent strains to maintain immunity against attacks by T6SS-active bacterial neighbors. The dual expression profile allows transcription of the three genes encoding immunity proteins independently of other T6SS proteins encoded within the same operon. One of these immunity proteins, TsiV2, protects against the T6SS effector VasX which is encoded immediately upstream of tsiV2. VasX is a secreted, lipid-binding protein that we previously characterized with respect to T6SS-mediated virulence towards the social amoeba Dictyostelium discoideum. Our data suggest the presence of an internal promoter in the open reading frame of vasX that drives expression of the downstream gene tsiV2. Furthermore, VasX is shown to act in conjunction with VasW, an accessory protein to VasX, to compromise the inner membrane of prokaryotic target cells. The dual regulatory profile of the T6SS immunity protein-encoding genes tsiV1, tsiV2, and tsiV3 permits V. cholerae to tightly control T6SS gene expression while maintaining immunity to T6SS activity.
Zdroje
1. WilliamsSG, VarcoeLT, AttridgeSR, ManningPA (1996) Vibrio cholerae Hcp, a secreted protein coregulated with HlyA. Infect Immun 64: 283–289.
2. BallisterER, LaiAH, ZuckermannRN, ChengY, MougousJD (2008) In vitro self-assembly of tailorable nanotubes from a simple protein building block. Proc Natl Acad Sci U S A 105: 3733–3738.
3. SilvermanJM, AgnelloDM, ZhengH, AndrewsBT, LiM, et al. (2013) Haemolysin coregulated protein is an exported receptor and chaperone of type VI secretion substrates. Mol Cell 51: 584–593.
4. PukatzkiS, MaAT, RevelAT, SturtevantD, MekalanosJJ (2007) Type VI secretion system translocates a phage tail spike-like protein into target cells where it cross-links actin. Proc Natl Acad Sci U S A 104: 15508–15513.
5. PukatzkiS, MaAT, SturtevantD, KrastinsB, SarracinoD, et al. (2006) Identification of a conserved bacterial protein secretion system in Vibrio cholerae using the Dictyostelium host model system. Proc Natl Acad Sci U S A 103: 1528–1533.
6. LeimanPG, BaslerM, RamagopalUA, BonannoJB, SauderJM, et al. (2009) Type VI secretion apparatus and phage tail-associated protein complexes share a common evolutionary origin. Proc Natl Acad Sci U S A 106: 4154–4159.
7. BaslerM, MekalanosJJ (2012) Type 6 secretion dynamics within and between bacterial cells. Science 337: 815.
8. BaslerM, PilhoferM, HendersonGP, JensenGJ, MekalanosJJ (2012) Type VI secretion requires a dynamic contractile phage tail-like structure. Nature 483: 182–186.
9. BonemannG, PietrosiukA, DiemandA, ZentgrafH, MogkA (2009) Remodelling of VipA/VipB tubules by ClpV-mediated threading is crucial for type VI protein secretion. EMBO J 28: 315–325.
10. MaAT, McAuleyS, PukatzkiS, MekalanosJJ (2009) Translocation of a Vibrio cholerae type VI secretion effector requires bacterial endocytosis by host cells. Cell Host Microbe 5: 234–243.
11. DongTG, HoBT, Yoder-HimesDR, MekalanosJJ (2013) Identification of T6SS-dependent effector and immunity proteins by Tn-seq in Vibrio cholerae. Proc Natl Acad Sci U S A 110: 2623–2628.
12. BrooksTM, UnterwegerD, BachmannV, KostiukB, PukatzkiS (2013) Lytic activity of the Vibrio cholerae type VI secretion toxin VgrG-3 is inhibited by the antitoxin TsaB. J Biol Chem 288: 7618–7625.
13. MaLS, LinJS, LaiEM (2009) An IcmF family protein, ImpLM, is an integral inner membrane protein interacting with ImpKL, and its walker a motif is required for type VI secretion system-mediated Hcp secretion in Agrobacterium tumefaciens. J Bacteriol 191: 4316–4329.
14. ShneiderMM, ButhSA, HoBT, BaslerM, MekalanosJJ, et al. (2013) PAAR-repeat proteins sharpen and diversify the type VI secretion system spike. Nature 500: 350–353.
15. ZinnakaY, CarpenterCCJr (1972) An enterotoxin produced by noncholera Vibrios. Johns Hopkins Med J 131: 403–411.
16. MacIntyreDL, MiyataST, KitaokaM, PukatzkiS (2010) The Vibrio cholerae type VI secretion system displays antimicrobial properties. Proc Natl Acad Sci U S A 107: 19520–19524.
17. ZhengJ, ShinOS, CameronDE, MekalanosJJ (2010) Quorum sensing and a global regulator TsrA control expression of type VI secretion and virulence in Vibrio cholerae. Proc Natl Acad Sci U S A 107: 21128–21133.
18. KitaokaM, MiyataST, BrooksTM, UnterwegerD, PukatzkiS (2011) VasH is a transcriptional regulator of the type VI secretion system functional in endemic and pandemic Vibrio cholerae. J Bacteriol 193: 6471–6482.
19. BernardCS, BrunetYR, GavioliM, LloubesR, CascalesE (2011) Regulation of type VI secretion gene clusters by sigma54 and cognate enhancer binding proteins. J Bacteriol 193: 2158–2167.
20. DongTG, MekalanosJJ (2012) Characterization of the RpoN regulon reveals differential regulation of T6SS and new flagellar operons in Vibrio cholerae O37 strain V52. Nucleic Acids Res
21. MiyataST, KitaokaM, BrooksTM, McAuleySB, PukatzkiS (2011) Vibrio cholerae requires the type VI secretion system virulence factor VasX to kill Dictyostelium discoideum. Infect Immun 79: 2941–2949.
22. UnterwegerD, KitaokaM, MiyataST, BachmannV, BrooksTM, et al. (2012) Constitutive type VI secretion system expression gives Vibrio cholerae intra- and interspecific competitive advantages. PLoS One 7: e48320.
23. HoodRD, SinghP, HsuF, GuvenerT, CarlMA, et al. (2010) A type VI secretion system of Pseudomonas aeruginosa targets a toxin to bacteria. Cell Host Microbe 7: 25–37.
24. MougousJD, CuffME, RaunserS, ShenA, ZhouM, et al. (2006) A virulence locus of Pseudomonas aeruginosa encodes a protein secretion apparatus. Science 312: 1526–1530.
25. SchwarzS, WestTE, BoyerF, ChiangWC, CarlMA, et al. (2010) Burkholderia type VI secretion systems have distinct roles in eukaryotic and bacterial cell interactions. PLoS Pathog 6(8): e1001068.
26. MurdochSL, TrunkK, EnglishG, FritschMJ, PourkarimiE, et al. (2011) The opportunistic pathogen Serratia marcescens utilizes type VI secretion to target bacterial competitors. J Bacteriol 193: 6057–6069.
27. AubertDF, FlannaganRS, ValvanoMA (2008) A novel sensor kinase-response regulator hybrid controls biofilm formation and type VI secretion system activity in Burkholderia cenocepacia. Infect Immun 76: 1979–1991.
28. BurtnickMN, BrettPJ, HardingSV, NgugiSA, RibotWJ, et al. (2011) The cluster 1 type VI secretion system is a major virulence determinant in Burkholderia pseudomallei. Infect Immun 79: 1512–1525.
29. BurtnickMN, DeShazerD, NairV, GherardiniFC, BrettPJ (2010) Burkholderia mallei cluster 1 type VI secretion mutants exhibit growth and actin polymerization defects in RAW 264.7 murine macrophages. Infect Immun 78: 88–99.
30. SchellMA, UlrichRL, RibotWJ, BrueggemannEE, HinesHB, et al. (2007) Type VI secretion is a major virulence determinant in Burkholderia mallei. Mol Microbiol 64: 1466–1485.
31. WangX, WangQ, XiaoJ, LiuQ, WuH, et al. (2009) Edwardsiella tarda T6SS component evpP is regulated by esrB and iron, and plays essential roles in the invasion of fish. Fish Shellfish Immunol 27: 469–477.
32. SuarezG, SierraJC, ShaJ, WangS, ErovaTE, et al. (2008) Molecular characterization of a functional type VI secretion system from a clinical isolate of Aeromonas hydrophila. Microb Pathog 44: 344–361.
33. BaslerM, HoBT, MekalanosJJ (2013) Tit-for-tat: type VI secretion system counterattack during bacterial cell-cell interactions. Cell 152: 884–894.
34. IshikawaT, SabharwalD, BromsJ, MiltonDL, SjostedtA, et al. (2012) Pathoadaptive conditional regulation of the type VI secretion system in Vibrio cholerae O1 strains. Infect Immun 80: 575–584.
35. RussellAB, HoodRD, BuiNK, LeRouxM, VollmerW, et al. (2011) Type VI secretion delivers bacteriolytic effectors to target cells. Nature 475: 343–347.
36. RussellAB, SinghP, BrittnacherM, BuiNK, HoodRD, et al. (2012) A widespread bacterial type VI secretion effector superfamily identified using a heuristic approach. Cell Host Microbe 11: 538–549.
37. GratiaA (1925) Sur un remarquable exemple d'antagonism entre deux souches de colibacille. Comptes Rendus des Seances et Memoires de la Societe de Biologie 93: 1040–1041.
38. ChakKF, KuoWS, LuFM, JamesR (1991) Cloning and characterization of the ColE7 plasmid. J Gen Microbiol 137: 91–100.
39. CooperPC, JamesR (1984) Two new E colicins, E8 and E9, produced by a strain of Escherichia coli. J Gen Microbiol 130: 209–215.
40. SchallerK, NomuraM (1976) Colicin E2 is DNA endonuclease. Proc Natl Acad Sci U S A 73: 3989–3993.
41. SeniorBW, HollandIB (1971) Effect of colicin E3 upon the 30S ribosomal subunit of Escherichia coli. Proc Natl Acad Sci U S A 68: 959–963.
42. BowmanCM, DahlbergJE, IkemuraT, KoniskyJ, NomuraM (1971) Specific inactivation of 16S ribosomal RNA induced by colicin E3 in vivo. Proc Natl Acad Sci U S A 68: 964–968.
43. BowmanCM, SidikaroJ, NomuraM (1971) Specific inactivation of ribosomes by colicin E3 in vitro and mechanism of immunity in colicinogenic cells. Nat New Biol 234: 133–137.
44. BoonT (1972) Inactivation of ribosomes in vitro by colicin E 3 and its mechanism of action. Proc Natl Acad Sci U S A 69: 549–552.
45. SchallerK, HoltjeJV, BraunV (1982) Colicin M is an inhibitor of murein biosynthesis. J Bacteriol 152: 994–1000.
46. ElkinsP, BunkerA, CramerWA, StauffacherCV (1997) A mechanism for toxin insertion into membranes is suggested by the crystal structure of the channel-forming domain of colicin E1. Structure 5: 443–458.
47. VetterIR, ParkerMW, TuckerAD, LakeyJH, PattusF, et al. (1998) Crystal structure of a colicin N fragment suggests a model for toxicity. Structure 6: 863–874.
48. ReevesP (1968) Mode of action of colicins of types E1, E2, E3, and K. J Bacteriol 96: 1700–1703.
49. LuriaSE (1964) On the Mechanisms of Action of Colicins. Ann Inst Pasteur (Paris) 107: SUPPL: 67–73.
50. ZakharovSD, CramerWA (2002) Insertion intermediates of pore-forming colicins in membrane two-dimensional space. Biochimie 84: 465–475.
51. ZakharovSD, KotovaEA, AntonenkoYN, CramerWA (2004) On the role of lipid in colicin pore formation. Biochim Biophys Acta 1666: 239–249.
52. ScheinSJ, KaganBL, FinkelsteinA (1978) Colicin K acts by forming voltage-dependent channels in phospholipid bilayer membranes. Nature 276: 159–163.
53. FieldsKL, LuriaSE (1969) Effects of colicins E1 and K on cellular metabolism. J Bacteriol 97: 64–77.
54. FieldsKL, LuriaSE (1969) Effects of colicins E1 and K on transport systems. J Bacteriol 97: 57–63.
55. PhillipsSK, CramerWA (1973) Properties of the fluorescence probe response associated with the transmission mechanism of colicin E1. Biochemistry 12: 1170–1176.
56. CascalesE, BuchananSK, DucheD, KleanthousC, LloubesR, et al. (2007) Colicin biology. Microbiol Mol Biol Rev 71: 158–229.
57. EspessetD, DucheD, BatyD, GeliV (1996) The channel domain of colicin A is inhibited by its immunity protein through direct interaction in the Escherichia coli inner membrane. EMBO J 15: 2356–2364.
58. CameronDE, UrbachJM, MekalanosJJ (2008) A defined transposon mutant library and its use in identifying motility genes in Vibrio cholerae. Proc Natl Acad Sci U S A 105: 8736–8741.
59. MetcalfWW, JiangW, DanielsLL, KimSK, HaldimannA, et al. (1996) Conditionally replicative and conjugative plasmids carrying lacZ alpha for cloning, mutagenesis, and allele replacement in bacteria. Plasmid 35: 1–13.
60. GuzmanLM, BelinD, CarsonMJ, BeckwithJ (1995) Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol 177: 4121–4130.
61. WhiteAP, Allen-VercoeE, JonesBW, DeVinneyR, KayWW, et al. (2007) An efficient system for markerless gene replacement applicable in a wide variety of enterobacterial species. Can J Microbiol 53: 56–62.
62. HsiaoA, LiuZ, JoelssonA, ZhuJ (2006) Vibrio cholerae virulence regulator-coordinated evasion of host immunity. Proc Natl Acad Sci U S A 103: 14542–14547.
63. MiyataST, KitaokaM, WieteskaL, FrechC, ChenN, et al. (2010) The Vibrio cholerae type VI secretion system: evaluating its role in the human disease cholera. Front Microbiol 1: 117.
64. ZhengJ, HoB, MekalanosJJ (2011) Genetic analysis of anti-amoebae and anti-bacterial activities of the type VI secretion system in Vibrio cholerae. PLoS One 6: e23876.
65. Gode-PotratzCJ, McCarterLL (2011) Quorum sensing and silencing in Vibrio parahaemolyticus. J Bacteriol 193: 4224–4237.
66. MaL, ZhangY, YanX, GuoL, WangL, et al. (2012) Expression of the type VI secretion system 1 component Hcp1 is indirectly repressed by OpaR in Vibrio parahaemolyticus. ScientificWorldJournal 2012: 982140.
67. EspessetD, CordaY, CunninghamK, BenedettiH, LloubesR, et al. (1994) The colicin A pore-forming domain fused to mitochondrial intermembrane space sorting signals can be functionally inserted into the Escherichia coli plasma membrane by a mechanism that bypasses the Tol proteins. Mol Microbiol 13: 1121–1131.
68. AokiSK, PammaR, HerndayAD, BickhamJE, BraatenBA, et al. (2005) Contact-dependent inhibition of growth in Escherichia coli. Science 309: 1245–1248.
69. RakinA, BoolgakowaE, HeesemannJ (1996) Structural and functional organization of the Yersinia pestis bacteriocin pesticin gene cluster. Microbiology 142(Pt 12): 3415–3424.
70. AokiSK, WebbJS, BraatenBA, LowDA (2009) Contact-dependent growth inhibition causes reversible metabolic downregulation in Escherichia coli. J Bacteriol 191: 1777–1786.
71. HanKD, AhnDH, LeeSA, MinYH, KwonAR, et al. (2013) Identification of chromosomal HP0892-HP0893 toxin-antitoxin proteins in Helicobacter pylori and structural elucidation of their protein-protein interaction. J Biol Chem 288(8): 6004–13.
72. DouganG, SaulM, WarrenG, SherrattD (1978) A Functional Map of Plasmid ColE1. Molecular and General Genetics 158: 325–327.
73. AndreoliPM, OverbeekeN, VeltkampE, van EmbdenJD, NijkampHJ (1978) Genetic map of the bacteriocinogenic plasmid CLO DF13 derived by insertion of the transposon Tn901. Mol Gen Genet 160: 1–11.
74. MaAT, MekalanosJJ (2010) In vivo actin cross-linking induced by Vibrio cholerae type VI secretion system is associated with intestinal inflammation. Proc Natl Acad Sci U S A 107: 4365–4370.
75. MiyataST, BachmannV, PukatzkiS (2013) Type VI secretion system regulation as a consequence of evolutionary pressure. Journal of Medical Microbiology 62(Pt 5): 663–76.
76. YangCC, KoniskyJ (1984) Colicin V-treated Escherichia coli does not generate membrane potential. J Bacteriol 158: 757–759.
77. DankertJR, UrataniY, GrabauC, CramerWA, HermodsonM (1982) On a domain structure of colicin E1. A COOH-terminal peptide fragment active in membrane depolarization. J Biol Chem 257: 3857–3863.
78. UrataniY, HoshinoT (1984) Pyocin R1 inhibits active transport in Pseudomonas aeruginosa and depolarizes membrane potential. J Bacteriol 157: 632–636.
79. JacobF, SiminovitchL, WollmanE (1952) Sur la biosynthese d'une colicine et sur son mode d'action. Annales de l'Institut Pasteur (Paris) 83: 295–315.
80. JakesKS, ModelP (1979) Mechanism of export of colicin E1 and colicin E3. J Bacteriol 138: 770–778.
81. CavardD, HowardSP, LloubesR, LazdunskiC (1989) High-level expression of the colicin A lysis protein. Mol Gen Genet 217: 511–519.
82. LuirinkJ, de GraafFK, OudegaB (1987) Uncoupling of synthesis and release of cloacin DF13 and its immunity protein by Escherichia coli. Mol Gen Genet 206: 126–132.
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2013 Číslo 12
- Očkování proti virové hemoragické horečce Ebola experimentální vakcínou rVSVDG-ZEBOV-GP
- Parazitičtí červi v terapii Crohnovy choroby a dalších zánětlivých autoimunitních onemocnění
- Koronavirus hýbe světem: Víte jak se chránit a jak postupovat v případě podezření?
Najčítanejšie v tomto čísle
- Influence of Mast Cells on Dengue Protective Immunity and Immune Pathology
- Myeloid Dendritic Cells Induce HIV-1 Latency in Non-proliferating CD4 T Cells
- Host Defense via Symbiosis in
- Coronaviruses as DNA Wannabes: A New Model for the Regulation of RNA Virus Replication Fidelity