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A competence-regulated toxin-antitoxin system in Haemophilus influenzae


Autoři: Hailey Findlay Black aff001;  Scott Mastromatteo aff001;  Sunita Sinha aff002;  Rachel L. Ehrlich aff003;  Corey Nislow aff004;  Joshua Chang Mell aff003;  Rosemary J. Redfield aff001
Působiště autorů: Department of Zoology, University of British Columbia, Vancouver, British Columbia, Canada aff001;  Sequencing + Bioinformatics Consortium, Office of the Vice-President, University of British Columbia, Vancouver, British Columbia, Canada aff002;  Department of Microbiology & Immunology, Center for Genomic Sciences, Drexel University College of Medicine, Philadelphia, Pennsylvania, United States of America aff003;  Department of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Columbia, Canada aff004
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0217255

Souhrn

Natural competence allows bacteria to respond to environmental and nutritional cues by taking up free DNA from their surroundings, thus gaining both nutrients and genetic information. In the Gram-negative bacterium Haemophilus influenzae, the genes needed for DNA uptake are induced by the CRP and Sxy transcription factors in response to lack of preferred carbon sources and nucleotide precursors. Here we show that one of these genes, HI0659, encodes the antitoxin of a competence-regulated toxin-antitoxin operon (‘toxTA’), likely acquired by horizontal gene transfer from a Streptococcus species. Deletion of the putative toxin (HI0660) restores uptake to the antitoxin mutant. The full toxTA operon was present in only 17 of the 181 strains we examined; complete deletion was seen in 22 strains and deletions removing parts of the toxin gene in 142 others. In addition to the expected Sxy- and CRP-dependent-competence promoter, HI0659/660 transcript analysis using RNA-seq identified an internal antitoxin-repressed promoter whose transcription starts within toxT and will yield nonfunctional protein. We propose that the most likely effect of unopposed toxin expression is non-specific cleavage of mRNAs and arrest or death of competent cells in the culture. Although the high frequency of toxT and toxTA deletions suggests that this competence-regulated toxin-antitoxin system may be mildly deleterious, it could also facilitate downregulation of protein synthesis and recycling of nucleotides under starvation conditions. Although our analyses were focused on the effects of toxTA, the RNA-seq dataset will be a useful resource for further investigations into competence regulation.

Klíčová slova:

Antitoxins – DNA – Gene regulation – Haemophilus influenzae – Operons – RNA analysis – Sequence alignment – Toxins


Zdroje

1. Harms A, Brodersen DE, Mitarai N, Gerdes K. Toxins, targets, and triggers: an overview of toxin-antitoxin biology. Molecular cell. 2018;70(5):768–84. doi: 10.1016/j.molcel.2018.01.003 29398446

2. Chan WT, Espinosa M, Yeo CC. Keeping the wolves at bay: antitoxins of prokaryotic type II toxin-antitoxin systems. Frontiers in Molecular Biosciences. 2016;3:9. doi: 10.3389/fmolb.2016.00009 27047942

3. Hall AM, Gollan B, Helaine S. Toxin–antitoxin systems: reversible toxicity. Current Opinion in Microbiology. 2017;36:102–10. doi: 10.1016/j.mib.2017.02.003 28279904

4. Ambur OH, Engelstädter J, Johnsen PJ, Miller EL, Rozen DE. Steady at the wheel: conservative sex and the benefits of bacterial transformation. Philos T R Soc B. 2016;371(1706): doi: 10.1098/rstb.2015.0528 27619692

5. Johnston C, Martin B, Fichant G, Polard P, Claverys J-P. Bacterial transformation: distribution, shared mechanisms and divergent control. Nat Rev Microbiol. 2014;12: 181–196. doi: 10.1038/nrmicro3199 24509783

6. Mell JC, Redfield RJ. Natural competence and the evolution of DNA uptake specificity. J Bacteriol. 2014;196(8):1471–1483. doi: 10.1128/JB.01293-13 24488316

7. Redfield RJ, Cameron ADS, Qian Q, Hinds J, Ali TR, Kroll JS, et al. A novel CRP-dependent regulon controls expression of competence genes in Haemophilus influenzae. J Mol Biol. 2005;4(8):735–747.

8. Sinha S, Mell JC, Redfield RJ. Seventeen Sxy-dependent cyclic AMP receptor protein site-regulated genes are needed for natural transformation in Haemophilus influenzae. J Bacteriol. 2012;194(19):5245–5254. doi: 10.1128/JB.00671-12 22821979

9. Macfadyen LP. Chen D, Vo HC, Liao D, Sinotte R, Redfield RJ. Competence development by Haemophilus influenzae is regulated by the availability of nucleic acid precursors. Mol Microbiol. 2001;40(3):700–707. doi: 10.1046/j.1365-2958.2001.02419.x 11359575

10. Sinha S, Mell JC, Redfield R. The availability of purine nucleotides regulates natural competence by controlling translation of the competence activator Sxy. Mol Microbiol. 2013;88(6):1106–1119. doi: 10.1111/mmi.12245 23663205

11. Cameron ADS, Redfield RJ. Non-canonical CRP sites control competence regulons in Escherichia coli and many other gamma-proteobacteria. Nucleic Acids Res. 2006;34(20): 6001–6014. doi: 10.1093/nar/gkl734 17068078

12. Jorgensen MG, Pandey DP, Jaskolska M, Gerdes J. HicA of Escherichia coli defines a novel family of translation-independent mRNA interferases in bacteria and archaea. J Bacteriol. 2009;191(4):1191–1199. doi: 10.1128/JB.01013-08 19060138

13. Christensen-Dalsgaard M, Gerdes K. Two higBA loci in the Vibrio cholerae superintegron encode mRNA cleaving enzymes and can stabilize plasmids. Mol Microbiol. 2006;62:397–411. doi: 10.1111/j.1365-2958.2006.05385.x 17020579

14. Makarova KS, Wolf YI, Koonin EV. Comprehensive comparative-genomic analysis of Type 2 toxin-antitoxin systems and related mobile stress elements in prokaryotes. Biology Direct. 2009;4(19): doi: 10.1186/1745-6150-4-19 19493340

15. Kelley LA, Mezulis S, Yates CM, Wass MN, Sternberg MJ. The Phyre2 web portal for protein modeling, prediction and analysis. Nature Protocols. 2015;10(6):845. doi: 10.1038/nprot.2015.053 25950237

16. Letunic I. Interactive Tree Of Life (iTOL): an online tool for phylogenetic tree display and annotation. Bioinformatics. 2007;23(1):127–128. doi: 10.1093/bioinformatics/btl529 17050570

17. Maughan H, Redfield RJ. Extensive variation in natural competence in Haemophilus influenzae. Evolution. 2009;63(7):1852–1866. doi: 10.1111/j.1558-5646.2009.00658.x 19239488

18. Dorocicz IR, Williams PM, Redfield RJ. The Haemophilus influenzae adenylate cyclase gene: cloning, sequence, and essential role in competence. J Bacteriol. 1993;175(22):7142–7149. doi: 10.1128/jb.175.22.7142-7149.1993 8226661

19. Page R, Peti W. Toxin-antitoxin systems in bacterial growth arrest and persistence. Nature Chemical Biology. 2016;12:208–214. doi: 10.1038/nchembio.2044 26991085

20. Williams PM, Bannister LA, Redfield RJ. The Haemophilus influenzae sxy-1 mutation is in a newly identified gene essential for competence. J Bacteriol. 1994;176(22):6789–6794. doi: 10.1128/jb.176.22.6789-6794.1994 7961436

21. Ma C, Redfield RJ. Point Mutations in a Peptidoglycan Biosynthesis Gene Cause Competence Induction in Haemophilus influenzae. J Bacteriol. 2000; 182(12):3323–3330. doi: 10.1128/jb.182.12.3323-3330.2000 10852860

22. Overgaard M, Borch J, Jorgensen MG, Gerdes K. Messenger RNA interferase RelE controls relBE transcription by conditional cooperativity. Mol Microbiol. 2008;69(4):841–857. doi: 10.1111/j.1365-2958.2008.06313.x 18532983

23. Turnbull KJ, Gerdes K. HicA toxin of Escherichia coli derepresses hicAB transcription to selectively produce HicB antitoxin. Mol Microbiol. 2017;104(5): 781–792. doi: 10.1111/mmi.13662 28266056

24. Sinha S, Cameron ADS, Redfield RJ. Sxy induces a CRP-S regulon in Escherichia coli. J Bacteriol. 2009;191(16):5180–5195. doi: 10.1128/JB.00476-09 19502395

25. Goeders N, Van Melderen L. Toxin-antitoxin systems as multilevel interaction systems. Toxins. 2014;6(1):304–324. doi: 10.3390/toxins6010304 24434905

26. Gmuender H, Kuratli K, Di Padova K, Gray CP, Keck W, Evers S. Gene expression changes triggered by exposure of Haemophilus influenzae to novobiocin or ciprofloxacin: combined transcription and translation analysis. Genome Res. 2001;11: 28–42. 11156613

27. Sweetman WA, Moxon ER, Bayliss CD. Induction of the SOS regulon of Haemophilus influenzae does not affect phase variation rates at tetranucleotide or dinucleotide repeats. Microbiology. 2005;151(8):2751–63.

28. Hurley JM, Cruz JW, Ouyang M, Woychik NA. Bacterial toxin RelE mediates frequent codon-independent mRNA cleavage from the 5′ end of coding regions in vivo. J Biol Chem. 2011;286:14770–14778. doi: 10.1074/jbc.M110.108969 21324908

29. Goeders N, Dreze P-L, Van Melderen L. Relaxed cleavage specificity within the RelE toxin family. J Bacteriol. 2013;195(11): 2541–2549. doi: 10.1128/JB.02266-12 23543711

30. Tollervey D. Molecular Biology: RNA lost in translation. Nature. 2006;440: 425–426. 16554791

31. Pandey DP, Gerdes K. Toxin–antitoxin loci are highly abundant in free-living but lost from host-associated prokaryotes. Nucleic Acids Res. 2005;33(3):966–976. doi: 10.1093/nar/gki201 15718296

32. Gordon GC, Cameron JC, Pfleger BF. RNA sequencing identifies new RNase III cleavage sites in Escherichia coli and reveals increased regulation of mRNA. mBio. 2017;8(2):e00128–17. doi: 10.1128/mBio.00128-17 28351917

33. Masuda H, Inouye M. Toxins of prokaryotic toxin-antitoxin systems with sequence-specific endoribonuclease activity. Toxins. 2017;9(4):140.

34. Audoux J, Philippe N, Chikhi R, Salson M, Gallopin M, Gabriel M, et al. DE-kupl: exhaustive capture of biological variation in RNA-seq data through k-mer decomposition. Genome Biology. 2017;18(1):243. doi: 10.1186/s13059-017-1372-2 29284518

35. Pedersen K, Zavialov AV, Pavlov MY, Elf J, Gerdes K, Ehrenberg M. The Bacterial Toxin RelE Displays Codon-Specific Cleavage of mRNAs in the Ribosomal A Site. Cell. 2003;112(1):133–140.

36. Neubauer C, Gao Y-G, Andersen KR, Dunham CM, Kelley AC, Hentschel J, et al. The Structural Basis for mRNA Recognition and Cleavage by the Ribosome-Dependent Endonuclease RelE. Cell. 2009;139(6):1084–1095. doi: 10.1016/j.cell.2009.11.015 20005802

37. Ramisetty BC, Santhosh RS. Endoribonuclease type II toxin–antitoxin systems: functional or selfish?. Microbiology. 2017;163(7):931–9. doi: 10.1099/mic.0.000487 28691660

38. Alexander HE, and Leidy G. Determination of inherited traits of H. influenzae by desoxyribonucleic acid fractions isolated from type-specific cells. J Exp Med. 1951;93:345–359. doi: 10.1084/jem.93.4.345 14824407

39. Chandler MS. The gene encoding cAMP receptor protein is required for competence development in Haemophilus influenzae Rd. P Natl Acad Sci USA. 1992;89(5):1626–30.

40. Blackall PJ, Klaasen HL, van den Bosch H, Kuhnert P, Frey J. Proposal of a new serovar of Actinobacillus pleuropneumoniae: serovar 15. Vet Microbiol. 2002;84(1–2):47–52. doi: 10.1016/s0378-1135(01)00428-x 11731158

41. Tracy E, Ye F, Baker BD, Munson RS. Construction of non-polar mutants in Haemophilus influenzae using FLP recombinase technology. BMC Mol Biol. 2008;9: doi: 10.1186/1471-2180-11-208

42. Herriott RM, Meyer EY, Vogt M, Modan M. Defined medium for growth of Haemophilus influenzae. J Bacteriol. 1970;101(2):513–6. 5308770

43. Poje G, Redfield RJ. Transformation of Haemophilus influenzae. Methods Mol Med. 2003;71:57–70. doi: 10.1385/1-59259-321-6:57 12374031

44. Barcak GJ, Chandler MS, Redfield RJ, Tomb JF. Genetic systems in Haemophilus influenzae. Method Enzymol. 1991 204:321–342.

45. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic Local Alignment Search Tool. J Mol Biol. 1990;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2 2231712

46. Katoh S. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol. 2013;30:772–780. doi: 10.1093/molbev/mst010 23329690

47. Maddison WP, Maddison DR. Mesquite: a modular system for evolutionary analysis. Version 3.02. 2013; http://mesquiteproject.org

48. Stamatakis A. RAxML Version 8: A tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics. 2014;30(9):1312–1313. doi: 10.1093/bioinformatics/btu033 24451623

49. Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics. 2014;30(14):2068–2069. doi: 10.1093/bioinformatics/btu153 24642063

50. Page AJ, Cummins CA, Hunt M, Wong VK, Reuter S, Holden MTG, et al. Roary: rapid large-scale prokaryote pan genome analysis. Bioinformatics. 2015;31(22):3691–3693. doi: 10.1093/bioinformatics/btv421 26198102

51. Charif D, Lobry JR. SeqinR 1.0–2: A contributed package to the R Project for statistical computing devoted to biological sequences retrieval and analysis. Structural Approaches to Sequence Evolution. Springer, Berlin, Heidelberg. 2007; https://doi.org/10.1007/978-3-540-35306-5_10

52. Li H, and Durbin R. Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics. 2009;5(14):1754–1760.

53. Love M, Anders S, Huber W. Differential analysis of RNA-Seq data at the gene level using the DESeq2 package. Heidelberg: European Molecular Biology Laboratory (EMBL). 2013.

54. Benjamini Y, and Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc. 1995;57(1):285–300.


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