A new Vibrio cholerae sRNA modulates colonization and affects release of outer membrane vesicles

Abstract

Summary We discovered a new small non‐coding RNA (sRNA) gene, vrrA of Vibrio cholerae O1 strain A1552. A vrrA mutant overproduces OmpA porin, and we demonstrate that the 140 nt VrrA RNA represses ompA translation by base‐pairing with the 5′ region of the mRNA. The RNA chaperone Hfq is not stringently required for VrrA action, but expression of the vrrA gene requires the membrane stress sigma factor, σ E , suggesting that VrrA acts on ompA in response to periplasmic protein folding stress. We also observed that OmpA levels inversely correlated with the number of outer membrane vesicles (OMVs), and that VrrA increased OMV production comparable to loss of OmpA. VrrA is the first sRNA known to control OMV formation. Moreover, a vrrA mutant showed a fivefold increased ability to colonize the intestines of infant mice as compared with the wild type. There was increased expression of the main colonization factor of V. cholerae , the toxin co‐regulated pili, in the vrrA mutant as monitored by immunoblot detection of the TcpA protein. VrrA overproduction caused a distinct reduction in the TcpA protein level. Our findings suggest that VrrA contributes to bacterial fitness in certain stressful environments, and modulates infection of the host intestinal tract.

Shown for reading. Its publisher allows quoting it, not republishing it.

References 60

1. 1.

   Balsalobre C, Silvan JM, Berglund S, Mizunoe Y, Uhlin BE, Wai SN. Release of the type I secreted alpha-haemolysin via outer membrane vesicles from Escherichia coli. Mol Microbiol. 2006;59:99-112.

   Not in our library10.1111/j.1365-2958.2005.04938.x

2. 2.

   Beveridge TJ. Structures of gram-negative cell walls and their derived membrane vesicles. J Bacteriol. 1999;181:4725-4733.

   Not in our library10.1128/jb.181.16.4725-4733.1999

3. 3.

   Brennan RG, Link TM. Hfq structure, function and ligand binding. Curr Opin Microbiol. 2007;10:125-133.

   Not in our library10.1016/j.mib.2007.03.015

4. 4.

   Camilli A, Mekalanos JJ. Use of recombinase gene fusions to identify Vibrio cholerae genes induced during infection. Mol Microbiol. 1995;18:671-683.

   Not in our library10.1111/j.1365-2958.1995.mmi_18040671.x

5. 5.

   Davis BM, Waldor MK. RNase E-dependent processing stabilizes MicX, a Vibrio cholerae sRNA. Mol Microbiol. 2007;65:373-385.

   Not in our library10.1111/j.1365-2958.2007.05796.x

6. 6.

   Davis BM, Quinones M, Pratt J, Ding Y, Waldor MK. Characterization of the small untranslated RNA RyhB and its regulon in Vibrio cholerae. J Bacteriol. 2005;187:4005-4014.

   Not in our library10.1128/jb.187.12.4005-4014.2005

7. 7.

   De Las Penas A, Connolly L, Gross CA. The sigmaE-mediated response to extracytoplasmic stress in Escherichia coli is transduced by RseA and RseB, two negative regulators of sigmaE. Mol Microbiol. 1997;24:373-385.

   Not in our library10.1046/j.1365-2958.1997.3611718.x

8. 8.

   Delcour AH. Structure and function of pore-forming beta-barrels from bacteria. J Mol Microbiol Biotechnol. 2002;4:1-10.

   Not in our library

9. 9.

   Ding Y, Davis BM, Waldor MK. Hfq is essential for Vibrio cholerae virulence and downregulates sigma expression. Mol Microbiol. 2004;53:345-354.

   Not in our library10.1111/j.1365-2958.2004.04142.x

10. 10.

    Douchin V, Bohn C, Bouloc P. Down-regulation of porins by a small RNA bypasses the essentiality of the regulated intramembrane proteolysis protease RseP in Escherichia coli. J Biol Chem. 2006;281:12253-12259.

    Not in our library10.1074/jbc.m600819200

11. 11.

    Faruque SM, Albert MJ, Mekalanos JJ. Epidemiology, genetics, and ecology of toxigenic Vibrio cholerae. Microbiol Mol Biol Rev. 1998;62:1301-1314.

    Not in our library10.1128/mmbr.62.4.1301-1314.1998

12. 12.

    Figueroa-Bossi N, Lemire S, Maloriol D, Balbontin R, Casadesus J, Bossi L. Loss of Hfq activates the sigmaE-dependent envelope stress response in Salmonella enterica. Mol Microbiol. 2006;62:838-852.

    Not in our library10.1111/j.1365-2958.2006.05413.x

13. 13.

    Furste JP, Pansegrau W, Frank R, Blocker H, Scholz P, Bagdasarian M, Lanka E. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene. 1986;48:119-131.

    Not in our library10.1016/0378-1119(86)90358-6

14. 14.

    Gardel CL, Mekalanos JJ. Alterations in Vibrio cholerae motility phenotypes correlate with changes in virulence factor expression. Infect Immun. 1996;64:2246-2255.

    Not in our library10.1128/iai.64.6.2246-2255.1996

15. 15.

    Hartz D, McPheeters DS, Traut R, Gold L. Extension inhibition analysis of translation initiation complexes. Meth Enzymol. 1988;164:419-425.

    Not in our library10.1016/s0076-6879(88)64058-4

16. 16.

    Hershberg R, Altuvia S, Margalit H. A survey of small RNA-encoding genes in Escherichia coli. Nucleic Acids Res. 2003;31:1813-1820.

    Not in our library10.1093/nar/gkg297

17. 17.

    Iwanaga M, Kuyyakanond T. Large production of cholera toxin by Vibrio cholerae O1 in yeast extract peptone water. J Clin Microbiol. 1987;25:2314-2316.

    Not in our library10.1128/jcm.25.12.2314-2316.1987

18. 18.

    Johansen J, Rasmussen AA, Overgaard M, Valentin-Hansen P. Conserved small non-coding RNAs that belong to the sigmaE regulon: role in down-regulation of outer membrane proteins. J Mol Biol. 2006;364:1-8.

    Not in our library10.1016/j.jmb.2006.09.004

19. 19.

    Kondo K, Takade A, Amako K. Release of the outer membrane vesicles from Vibrio cholerae and Vibrio parahaemolyticus. Microbiol Immunol. 1993;37:149-152.

    Not in our library10.1111/j.1348-0421.1993.tb03192.x

20. 20.

    Kouokam JC, Wai SN, Fallman M, Dobrindt U, Hacker J, Uhlin BE. Active cytotoxic necrotizing factor 1 associated with outer membrane vesicles from uropathogenic Escherichia coli. Infect Immun. 2006;74:2022-2030.

    Not in our library10.1128/iai.74.4.2022-2030.2006

21. 21.

    Kuehn MJ, Kesty NC. Bacterial outer membrane vesicles and the host–pathogen interaction. Genes Dev. 2005;19:2645-2655.

    Not in our library10.1101/gad.1299905

22. 22.

    Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680-685.

    Not in our library10.1038/227680a0

23. 23.

    Lee SH, Hava DL, Waldor MK, Camilli A. Regulation and temporal expression patterns of Vibrio cholerae virulence genes during infection. Cell. 1999;99:625-634.

    Not in our library10.1016/s0092-8674(00)81551-2

24. 24.

    Lenz DH, Mok KC, Lilley BN, Kulkarni RV, Wingreen NS, Bassler BL. The small RNA chaperone Hfq and multiple small RNAs control quorum sensing in Vibrio harveyi and Vibrio cholerae. Cell. 2004;118:69-82.

    Not in our library10.1016/j.cell.2004.06.009

25. 25.

    Lenz DH, Miller MB, Zhu J, Kulkarni RV, Bassler BL. CsrA and three redundant small RNAs regulate quorum sensing in Vibrio cholerae. Mol Microbiol. 2005;58:1186-1202.

    Not in our library10.1111/j.1365-2958.2005.04902.x

26. 26.

    Livny J, Fogel MA, Davis BM, Waldor MK. sRNAPredict: an integrative computational approach to identify sRNAs in bacterial genomes. Nucleic Acids Res. 2005;33:4096-4105.

    Not in our library10.1093/nar/gki715

27. 27.

    Lutz R, Bujard H. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res. 1997;25:1203-1210.

    Not in our library10.1093/nar/25.6.1203

28. 28.
    Release of outer membrane vesicles by Gram‐negative bacteria is a novel envelope stress response · 2006

    No notice
29. 29.

    Majdalani N, Vanderpool CK, Gottesman S. Bacterial small RNA regulators. Crit Rev Biochem Mol Biol. 2005;40:93-113.

    Not in our library10.1080/10409230590918702

30. 30.

    Mashburn-Warren LM, Whiteley M. Special delivery: vesicle trafficking in prokaryotes. Mol Microbiol. 2006;61:839-846.

    Not in our library10.1111/j.1365-2958.2006.05272.x

31. 31.

    Mey AR, Craig SA, Payne SM. Characterization of Vibrio cholerae RyhB: the RyhB regulon and role of ryhB in biofilm formation. Infect Immun. 2005;73:5706-5719.

    Not in our library10.1128/iai.73.9.5706-5719.2005

32. 32.
    σ E ‐dependent small RNAs of Salmonella respond to membrane stress by accelerating global omp mRNA decay · 2006

    No notice
33. 33.

    Rehmsmeier M, Steffen P, Hochsmann M, Giegerich R. Fast and effective prediction of microRNA/target duplexes. RNA. 2004;10:1507-1517.

    Not in our library10.1261/rna.5248604

34. 34.
    Conserved and Variable Functions of the σE Stress Response in Related Genomes · 2005

    No notice
35. 35.

    Romby P, Vandenesch F, Wagner EG. The role of RNAs in the regulation of virulence-gene expression. Curr Opin Microbiol. 2006;9:229-236.

    Not in our library10.1016/j.mib.2006.02.005

36. 36.

    Schild S, Tamayo R, Nelson EJ, Qadri F, Calderwood SB, Camilli A. Genes induced late in infection increase fitness of Vibrio cholerae after release into the environment. Cell Host Microbe. 2007;2:264-277.

    Not in our library10.1016/j.chom.2007.09.004

37. 37.

    Schooling SR, Beveridge TJ. Membrane vesicles: an overlooked component of the matrices of biofilms. J Bacteriol. 2006;188:5945-5957.

    Not in our library10.1128/jb.00257-06

38. 38.

    Sharma CM, Darfeuille F, Plantinga TH, Vogel J. A small RNA regulates multiple ABC transporter mRNAs by targeting C/A-rich elements inside and upstream of ribosome-binding sites. Genes Dev. 2007;21:2804-2817.

    Not in our library10.1101/gad.447207

39. 39.

    Sheehan B, Klarsfeld A, Msadek T, Cossart P. Differential activation of virulence gene expression by PrfA, the Listeria monocytogenes virulence regulator. J Bacteriol. 1995;177:6469-6476.

    Not in our library10.1128/jb.177.22.6469-6476.1995

40. 40.

    Skovierova H, Rowley G, Rezuchova B, Homerova D, Lewis C, Roberts M, Kormanec J. Identification of the sigmaE regulon of Salmonella enterica serovar Typhimurium. Microbiology. 2006;152:1347-1359.

    Not in our library10.1099/mic.0.28744-0

Cited by papers we hold 9

• Cracking Open Bacterial Membrane Vesicles · 2020
  No notice
• Bacterial Membrane Vesicles as Mediators of Microbe – Microbe and Microbe – Host Community Interactions · 2020
  No notice
• Extracellular membrane vesicles in the three domains of life and beyond · 2018
  No notice
• Membrane Distribution of the Pseudomonas Quinolone Signal Modulates Outer Membrane Vesicle Production in Pseudomonas aeruginosa · 2017
  No notice
• A novel mechanism for the biogenesis of outer membrane vesicles in Gram-negative bacteria · 2016
  No notice
• Identification of a Novel Small Non-Coding RNA Modulating the Intracellular Survival of Brucella melitensis · 2015
  Not checked
• Outer Membrane Vesicle-Mediated Export of Processed PrtV Protease from Vibrio cholerae. · 2015
  Not checked
• Vibrio cholerae utilizes direct sRNA regulation in expression of a biofilm matrix protein. · 2014
  Not checked
• A Staphylococcus aureus Small RNA Is Required for Bacterial Virulence and Regulates the Expression of an Immune-Evasion Molecule · 2010
  No notice

Reads

A person counts once a day per paper. We keep no cookie and no address: a reader is a hash that changes every day. Any month under five reads shows no number.