A Small Non-Coding RNA Facilitates Bacterial Invasion and Intracellular Replication by Modulating the Expression of Virulence Factors

English version České info

Small non-coding RNAs (sRNAs) that act as regulators of gene expression have been identified in all kingdoms of life, including microRNA (miRNA) and small interfering RNA (siRNA) in eukaryotic cells. Numerous sRNAs identified in Salmonella are encoded by genes located at Salmonella pathogenicity islands (SPIs) that are commonly found in pathogenic strains. Whether these sRNAs are important for Salmonella pathogenesis and virulence in animals has not been reported. In this study, we provide the first direct evidence that a pathogenicity island-encoded sRNA, IsrM, is important for Salmonella invasion of epithelial cells, intracellular replication inside macrophages, and virulence and colonization in mice. IsrM RNA is expressed in vitro under conditions resembling those during infection in the gastrointestinal tract. Furthermore, IsrM is found to be differentially expressed in vivo, with higher expression in the ileum than in the spleen. IsrM targets the mRNAs coding for SopA, a SPI-1 effector, and HilE, a global regulator of the expression of SPI-1 proteins, which are major virulence factors essential for bacterial invasion. Mutations in IsrM result in disregulation of expression of HilE and SopA, as well as other SPI-1 genes whose expression is regulated by HilE. Salmonella with deletion of isrM is defective in bacteria invasion of epithelial cells and intracellular replication/survival in macrophages. Moreover, Salmonella with mutations in isrM is attenuated in killing animals and defective in growth in the ileum and spleen in mice. Our study has shown that IsrM sRNA functions as a pathogenicity island-encoded sRNA directly involved in Salmonella pathogenesis in animals. Our results also suggest that sRNAs may represent a distinct class of virulence factors that are important for bacterial infection in vivo.

Vyšlo v časopise: A Small Non-Coding RNA Facilitates Bacterial Invasion and Intracellular Replication by Modulating the Expression of Virulence Factors. PLoS Pathog 7(9): e32767. doi:10.1371/journal.ppat.1002120
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002120

Souhrn

Zdroje

1. OlsenSJMacKinnonLCGouldingJSBeanNHSlutskerL 2000 Surveillance for foodborne-disease outbreaks–United States, 1993–1997. MMWR CDC Surveill Summ 49 1 62

2. SchmidtHHenselM 2004 Pathogenicity islands in bacterial pathogenesis. Clin Microbiol Rev 17 14 56

3. GalanJEWolf-WatzH 2006 Protein delivery into eukaryotic cells by type III secretion machines. Nature 444 567 573

4. AbrahamsGLHenselM 2006 Manipulating cellular transport and immune responses: dynamic interactions between intracellular Salmonella enterica and its host cells. Cell Microbiol 8 728 737

5. GalanJE 2001 Salmonella interactions with host cells: type III secretion at work. Annu Rev Cell Dev Biol 17 53 86

6. WatermanSRHoldenDW 2003 Functions and effectors of the Salmonella pathogenicity island 2 type III secretion system. Cell Microbiol 5 501 511

7. HueckCJ 1998 Type III protein secretion systems in bacterial pathogens of animals and plants. Microbiol Mol Biol Rev 62 379 433

8. EllermeierJRSlauchJM 2007 Adaptation to the host environment: regulation of the SPI1 type III secretion system in Salmonella enterica serovar Typhimurium. Curr Opin Microbiol 10 24 29

9. GiacomodonatoMNUzzauSBacciuDCaccuriRSarnackiSH 2007 SipA, SopA, SopB, SopD and SopE2 effector proteins of Salmonella enterica serovar Typhimurium are synthesized at late stages of infection in mice. Microbiology 153 1221 1228

10. PfeiferCGMarcusSLSteele-MortimerOKnodlerLAFinlayBB 1999 Salmonella typhimurium virulence genes are induced upon bacterial invasion into phagocytic and nonphagocytic cells. Infect Immun 67 5690 5698

11. Steele-MortimerOBrumellJHKnodlerLAMeresseSLopezA 2002 The invasion-associated type III secretion system of Salmonella enterica serovar Typhimurium is necessary for intracellular proliferation and vacuole biogenesis in epithelial cells. Cell Microbiol 4 43 54

12. GongHSuJBaiYMiaoLKimK 2009 Characterization of the expression of Salmonella Type III secretion system factor PrgI, SipA, SipB, SopE2, SpaO, and SptP in cultures and in mice. BMC Microbiol 9 73

13. GongHVuGPBaiYYangELiuF 2010 Differential expression of Salmonella type III secretion system factors InvJ, PrgJ, SipC, SipD, SopA and SopB in cultures and in mice. Microbiology 156 116 127

14. HannonGJRivasFVMurchisonEPSteitzJA 2006 The expanding universe of noncoding RNAs. Cold Spring Harb Symp Quant Biol 71 551 564

15. StorzG 2002 An expanding universe of noncoding RNAs. Science 296 1260 1263

16. Toledo-AranaARepoilaFCossartP 2007 Small noncoding RNAs controlling pathogenesis. Curr Opin Microbiol 10 182 188

17. PapenfortKVogelJ 2010 Regulatory RNA in bacterial pathogens. Cell Host Microbe 8 116 127

18. StorzGOpdykeJAWassarmanKM 2006 Regulating bacterial transcription with small RNAs. Cold Spring Harb Symp Quant Biol 71 269 273

19. VogelJ 2009 A rough guide to the non-coding RNA world of Salmonella. Mol Microbiol 71 1 11

20. StorzGOpdykeJAZhangA 2004 Controlling mRNA stability and translation with small, noncoding RNAs. Curr Opin Microbiol 7 140 144

21. BabitzkePRomeoT 2007 CsrB sRNA family: sequestration of RNA-binding regulatory proteins. Curr Opin Microbiol 10 156 163

22. WassarmanKM 2007 6S RNA: a small RNA regulator of transcription. Curr Opin Microbiol 10 164 168

23. Padalon-BrauchGHershbergRElgrably-WeissMBaruchKRosenshineI 2008 Small RNAs encoded within genetic islands of Salmonella typhimurium show host-induced expression and role in virulence. Nucleic Acids Res 36 1913 1927

24. PfeifferVSittkaATomerRTedinKBrinkmannV 2007 A small non-coding RNA of the invasion gene island (SPI-1) represses outer membrane protein synthesis from the Salmonella core genome. Mol Microbiol 66 1174 1191

25. SittkaALucchiniSPapenfortKSharmaCMRolleK 2008 Deep sequencing analysis of small noncoding RNA and mRNA targets of the global post-transcriptional regulator, Hfq. PLoS Genet 4 e1000163

26. ArricauNHermantDWaxinHEcobichonCDuffeyPS 1998 The RcsB-RcsC regulatory system of Salmonella typhi differentially modulates the expression of invasion proteins, flagellin and Vi antigen in response to osmolarity. Mol Microbiol 29 835 850

27. GalanJECurtissR3rd 1990 Expression of Salmonella typhimurium genes required for invasion is regulated by changes in DNA supercoiling. Infect Immun 58 1879 1885

28. TarteraCMetcalfES 1993 Osmolarity and growth phase overlap in regulation of Salmonella typhi adherence to and invasion of human intestinal cells. Infect Immun 61 3084 3089

29. JonesBDFalkowS 1994 Identification and characterization of a Salmonella typhimurium oxygen-regulated gene required for bacterial internalization. Infect Immun 62 3745 3752

30. LeeCAFalkowS 1990 The ability of Salmonella to enter mammalian cells is affected by bacterial growth state. Proc Natl Acad Sci U S A 87 4304 4308

31. CummingsJHPomareEWBranchWJNaylorCPMacfarlaneGT 1987 Short chain fatty acids in human large intestine, portal, hepatic and venous blood. Gut 28 1221 1227

32. Van ImmerseelFDe BuckJDe SmetIPasmansFHaesebrouckF 2004 Interactions of butyric acid- and acetic acid-treated Salmonella with chicken primary cecal epithelial cells in vitro. Avian Dis 48 384 391

33. KrugerJRehmsmeierM 2006 RNAhybrid: microRNA target prediction easy, fast and flexible. Nucleic Acids Res 34 W451 454

34. TjadenBGoodwinSSOpdykeJAGuillierMFuDX 2006 Target prediction for small, noncoding RNAs in bacteria. Nucleic Acids Res 34 2791 2802

35. BaxterMAFahlenTFWilsonRLJonesBD 2003 HilE interacts with HilD and negatively regulates hilA transcription and expression of the Salmonella enterica serovar Typhimurium invasive phenotype. Infect Immun 71 1295 1305

36. UrbanJHVogelJ 2007 Translational control and target recognition by Escherichia coli small RNAs in vivo. Nucleic Acids Res 35 1018 1037

37. BuchmeierNAHeffronF 1989 Intracellular survival of wild-type Salmonella typhimurium and macrophage-sensitive mutants in diverse populations of macrophages. Infect Immun 57 1 7

38. EllermeierCDEllermeierJRSlauchJM 2005 HilD, HilC and RtsA constitute a feed forward loop that controls expression of the SPI1 type three secretion system regulator hilA in Salmonella enterica serovar Typhimurium. Mol Microbiol 57 691 705

39. ZhangYHigashideWMMcCormickBAChenJZhouD 2006 The inflammation-associated Salmonella SopA is a HECT-like E3 ubiquitin ligase. Mol Microbiol 62 786 793

40. DiaoJZhangYHuibregtseJMZhouDChenJ 2008 Crystal structure of SopA, a Salmonella effector protein mimicking a eukaryotic ubiquitin ligase. Nat Struct Mol Biol 15 65 70

41. HicksSWGalánJE 2010 Hijacking the host ubiquitin pathway: structural strategies of bacterial E3 ubiquitin ligases. Curr Opin Microbiol 13 41 46

42. RaffatelluMWilsonRPChessaDAndrews-PolymenisHTranQT 2005 SipA, SopA, SopB, SopD, and SopE2 contribute to Salmonella enterica serotype typhimurium invasion of epithelial cells. Infect Immun 73 146 154

43. BrennanRGLinkTM 2007 Hfq structure, function and ligand binding. Curr Opin Microbiol 10 125 133

44. Valentin-HansenPEriksenMUdesenC 2004 The bacterial Sm-like protein Hfq: a key player in RNA transactions. Mol Microbiol 51 1525 1533

45. CainRJHaywardRDKoronakisV 2008 Deciphering interplay between Salmonella invasion effectors. PLoS Pathog 4 e1000037

46. WinnenBSchlumbergerMCSturmASchupbachKSiebenmannS 2008 Hierarchical effector protein transport by the Salmonella Typhimurium SPI-1 type III secretion system. PLoS ONE 3 e2178

47. DatsenkoKAWannerBL 2000 One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97 6640 6645

48. LuSKilloranPBFangFCRileyLW 2002 The global regulator ArcA controls resistance to reactive nitrogen and oxygen intermediates in Salmonella enterica serovar Enteritidis. Infect Immun 70 451 461

49. SuJGongHLaiJMainALuS 2009 The potassium transporter Trk and external potassium modulate Salmonella enterica protein secretion and virulence. Infect Immun 77 667 675

50. LuSMangesARXuYFangFCRileyLW 1999 Analysis of virulence of clinical isolates of Salmonella enteritidis in vivo and in vitro. Infect Immun 67 5651 5657

51. McClellandMSandersonKESpiethJCliftonSWLatreilleP 2001 Complete genome sequence of Salmonella enterica serovar Typhimurium LT2. Nature 413 852 856