Shewanella oneidensis Hfq promotes exponential phase growth, stationary phase culture density, and cell survival

BMC Microbiology

Brett J Pellock¹  Jessica M Osborn¹  Katherine E Douglas¹  Taylor Mezoian¹  Zachary Sexton¹  Nicholas Q Mazzucca¹  Matthew T Goulet¹  Taylor M Hunt¹  Meghan L Keane¹  Christopher M Brennan¹ 

[1] Department of Biology, Providence College, Providence, RI, USA

关键词: Stationary phase survival;    Oxidative stress;    Metal reduction;    Hfq;    Shewanella oneidensis;   

Others  :  1144423
DOI  :  10.1186/1471-2180-13-33

 received in 2012-11-09, accepted in 2013-01-21,  发布年份 2013

【 摘 要 】

Background

Hfq is an RNA chaperone protein that has been broadly implicated in sRNA function in bacteria. Here we describe the construction and characterization of a null allele of the gene that encodes the RNA chaperone Hfq in Shewanella oneidensis strain MR-1, a dissimilatory metal reducing bacterium.

Results

Loss of hfq in S. oneidensis results in a variety of mutant phenotypes, all of which are fully complemented by addition of a plasmid-borne copy of the wild type hfq gene. Aerobic cultures of the hfq∆ mutant grow more slowly through exponential phase than wild type cultures, and hfq∆ cultures reach a terminal cell density in stationary phase that is ~2/3 of that observed in wild type cultures. We have observed a similar growth phenotype when the hfq∆ mutant is cultured under anaerobic conditions with fumarate as the terminal electron acceptor, and we have found that the hfq∆ mutant is defective in Cr(VI) reduction. Finally, the hfq∆ mutant exhibits a striking loss of colony forming units in extended stationary phase and is highly sensitive to oxidative stress induced by H₂O₂ or methyl viologen (paraquat).

Conclusions

The hfq mutant in S. oneidensis exhibits pleiotropic phenotypes, including a defect in metal reduction. Our results also suggest that hfq mutant phenotypes in S. oneidensis may be at least partially due to increased sensitivity to oxidative stress.

【 授权许可】

   
2013 Brennan et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Geissmann TA, Touati D: Hfq, a new chaperoning role: binding to messenger RNA determines access for small RNA regulator. EMBO J 2004, 23(2):396-405.
• [2]Gottesman S: The small RNA regulators of Escherichia coli: roles and mechanisms. Annu Rev Microbiol 2004, 58:303-328.
• [3]Moller T, Franch T, Hojrup P, Keene DR, Bachinger HP, Brennan RG, Valentin-Hansen P: Hfq: a bacterial Sm-like protein that mediates RNA-RNA interaction. Mol Cell 2002, 9(1):23-30.
• [4]Panja S, Woodson SA: Hexamer to monomer equilibrium of E. coli Hfq in solution and its impact on RNA annealing. J Mol Biol 2012, 417(5):406-412.
• [5]Tsui HC, Leung HC, Winkler ME: Characterization of broadly pleiotropic phenotypes caused by an hfq insertion mutation in Escherichia coli K-12. Mol Microbiol 1994, 13(1):35-49.
• [6]Sittka A, Pfeiffer V, Tedin K, Vogel J: The RNA chaperone Hfq is essential for the virulence of Salmonella typhimurium. Mol Microbiol 2007, 63(1):193-217.
• [7]Ding Y, Davis BM, Waldor MK: Hfq is essential for Vibrio cholerae virulence and downregulates sigma expression. Mol Microbiol 2004, 53(1):345-354.
• [8]Muffler A, Traulsen DD, Fischer D, Lange R, Hengge-Aronis R: The RNA-binding protein HF-I plays a global regulatory role which is largely, but not exclusively, due to its role in expression of the sigmaS subunit of RNA polymerase in Escherichia coli. J Bacteriol 1997, 179(1):297-300.
• [9]Myers CR, Nealson KH: Bacterial manganese reduction and growth with manganese oxide as the sole electron acceptor. Science 1988, 240(4857):1319-1321.
• [10]Nealson KH, Saffarini D: Iron and manganese in anaerobic respiration: environmental significance, physiology, and regulation. Annu Rev Microbiol 1994, 48:311-343.
• [11]Lovley DR: Bug juice: harvesting electricity with microorganisms. Nat Rev Microbiol 2006, 4(7):497-508.
• [12]Heidelberg JF, Paulsen IT, Nelson KE, Gaidos EJ, Nelson WC, Read TD, Eisen JA, Seshadri R, Ward N, Methe B, et al.: Genome sequence of the dissimilatory metal ion-reducing bacterium Shewanella oneidensis. Nat Biotechnol 2002, 20(11):1118-1123.
• [13]Becker A, Schmidt M, Jager W, Puhler A: New gentamicin-resistance and lacZ promoter-probe cassettes suitable for insertion mutagenesis and generation of transcriptional fusions. Gene 1995, 162(1):37-39.
• [14]Alting-Mees MA, Short JM: pBluescript II: gene mapping vectors. Nucleic Acids Res 1989, 17(22):9494.
• [15]Edwards RA, Keller LH, Schifferli DM: Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression. Gene 1998, 207(2):149-157.
• [16]Miller VL, Mekalanos JJ: A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J Bacteriol 1988, 170(6):2575-2583.
• [17]Tsui HC, Feng G, Winkler ME: Transcription of the mutL repair, miaA tRNA modification, hfq pleiotropic regulator, and hflA region protease genes of Escherichia coli K-12 from clustered Esigma32-specific promoters during heat shock. J Bacteriol 1996, 178(19):5719-5731.
• [18]Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM 2nd, Peterson KM: Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 1995, 166(1):175-176.
• [19]Simon R, Priefer U, Puhler A: A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria. Nat Biotech 1983, 1:784-791.
• [20]Zhang A, Wassarman KM, Ortega J, Steven AC, Storz G: The Sm-like Hfq protein increases OxyS RNA interaction with target mRNAs. Mol Cell 2002, 9(1):11-22.
• [21]Urone PF: Stability of colorimetric reagent for chromium, s-diphenylcarbazide, in various solvents. Anal Chem 1955, 27:1354-1355.
• [22]Dukan S, Nystrom T: Bacterial senescence: stasis results in increased and differential oxidation of cytoplasmic proteins leading to developmental induction of the heat shock regulon. Genes Dev 1998, 12(21):3431-3441.
• [23]Navarro Llorens JM, Tormo A, Martinez-Garcia E: Stationary phase in gram-negative bacteria. FEMS Microbiol Rev 2010, 34(4):476-495.
• [24]Geng J, Song Y, Yang L, Feng Y, Qiu Y, Li G, Guo J, Bi Y, Qu Y, Wang W, et al.: Involvement of the post-transcriptional regulator Hfq in Yersinia pestis virulence. PLoS One 2009, 4(7):e6213.
• [25]Guisbert E, Rhodius VA, Ahuja N, Witkin E, Gross CA: Hfq modulates the sigmaE-mediated envelope stress response and the sigma32-mediated cytoplasmic stress response in Escherichia coli. J Bacteriol 2007, 189(5):1963-1973.
• [26]Sonnleitner E, Schuster M, Sorger-Domenigg T, Greenberg EP, Blasi U: Hfq-dependent alterations of the transcriptome profile and effects on quorum sensing in Pseudomonas aeruginosa. Mol Microbiol 2006, 59(5):1542-1558.
• [27]Oliver JD: The viable but nonculturable state in bacteria. J Microbiol 2005, 43(Spec No):93-100.
• [28]Lease RA, Cusick ME, Belfort M: Riboregulation in Escherichia coli: DsrA RNA acts by RNA:RNA interactions at multiple loci. Proc Natl Acad Sci USA 1998, 95(21):12456-12461.
• [29]Majdalani N, Cunning C, Sledjeski D, Elliott T, Gottesman S: DsrA RNA regulates translation of RpoS message by an anti-antisense mechanism, independent of its action as an antisilencer of transcription. Proc Natl Acad Sci USA 1998, 95(21):12462-12467.
• [30]Majdalani N, Hernandez D, Gottesman S: Regulation and mode of action of the second small RNA activator of RpoS translation, RprA. Mol Microbiol 2002, 46(3):813-826.
• [31]Zhang A, Altuvia S, Tiwari A, Argaman L, Hengge-Aronis R, Storz G: The OxyS regulatory RNA represses rpoS translation and binds the Hfq (HF-I) protein. EMBO J 1998, 17(20):6061-6068.
• [32]Vogel J, Luisi BF: Hfq and its constellation of RNA. Nat Rev Microbiol 2011, 9(8):578-589.
• [33]Yang Y, McCue LA, Parsons AB, Feng S, Zhou J: The tricarboxylic acid cycle in Shewanella oneidensis is independent of Fur and RyhB control. BMC Microbiol 2010, 10:264. BioMed Central Full Text