1887

Abstract

In our previous study, extensive genomic rearrangements were found in two strains of the Gram-negative anaerobic bacterium () , and most of these rearrangements were associated with mobile genetic elements such as insertion sequences and conjugative transposons (CTns). CTnPg1, identified in strain ATCC 33277, was the first complete CTn reported for the genus In the present study, we found that CTnPg1 can be transferred from strain ATCC 33277 to another strain, W83, at a frequency of 10 to 10. The excision of CTnPg1 from the chromosome in a donor cell depends on an integrase (Int; PGN_0094) encoded in CTnPg1, whereas CTnPg1 excision is independent of PGN_0084 (a DNA topoisomerase I homologue; Exc) encoded within CTnPg1 and (PGN_1057) on the donor chromosome. Intriguingly, however, the transfer of CTnPg1 between strains requires RecA function in the recipient. Sequencing analysis of CTnPg1-integrated sites on the chromosomes of transconjugants revealed that the consensus attachment () sequence is a 13 bp sequence, TTTTCNNNNAAAA. We further report that CTnPg1 is able to transfer to two other bacterial species, and In addition, CTnPg1-like CTns are located in the genomes of other oral anaerobic bacteria, , and , with the same consensus sequence. These results suggest that CTns in the CTnPg1 family are widely distributed among oral anaerobic Gram-negative bacteria found in humans and play important roles in horizontal gene transfer among these bacteria.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.047803-0
2011-07-01
2019-10-22
Loading full text...

Full text loading...

/deliver/fulltext/micro/157/7/2022.html?itemId=/content/journal/micro/10.1099/mic.0.047803-0&mimeType=html&fmt=ahah

References

  1. Abaibou H. , Chen Z. , Olango G. J. , Liu Y. , Edwards J. , Fletcher H. M. . ( 2001; ). vimA gene downstream of recA is involved in virulence modulation in Porphyromonas gingivalis W83. . Infect Immun 69:, 325–335. [CrossRef].[PubMed].
    [Google Scholar]
  2. Altschul S. F. , Madden T. L. , Schäffer A. A. , Zhang J. , Zhang Z. , Miller W. , Lipman D. J. . ( 1997; ). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. . Nucleic Acids Res 25:, 3389–3402. [CrossRef].[PubMed].
    [Google Scholar]
  3. Bacic M. , Parker A. C. , Stagg J. , Whitley H. P. , Wells W. G. , Jacob L. A. , Smith C. J. . ( 2005; ). Genetic and structural analysis of the Bacteroides conjugative transposon CTn341. . J Bacteriol 187:, 2858–2869. [CrossRef].[PubMed].
    [Google Scholar]
  4. Beck J. , Garcia R. , Heiss G. , Vokonas P. S. , Offenbacher S. . ( 1996; ). Periodontal disease and cardiovascular disease. . J Periodontol 67: Suppl 1123–1137.[PubMed].[CrossRef]
    [Google Scholar]
  5. Bedzyk L. A. , Shoemaker N. B. , Young K. E. , Salyers A. A. . ( 1992; ). Insertion and excision of Bacteroides conjugative chromosomal elements. . J Bacteriol 174:, 166–172.[PubMed].
    [Google Scholar]
  6. Bellen H. J. , Levis R. W. , Liao G. , He Y. , Carlson J. W. , Tsang G. , Evans-Holm M. , Hiesinger P. R. , Schulze K. L. et al. ( 2004; ). The BDGP gene disruption project: single transposon insertions associated with 40% of Drosophila genes. . Genetics 167:, 761–781. [CrossRef].[PubMed].
    [Google Scholar]
  7. Chen T. , Yu W.-H. , Izard J. , Baranova O. V. , Lakshmanan A. , Dewhirst F. E. . ( 2010; ). The Human Oral Microbiome Database: a web accessible resource for investigating oral microbe taxonomic and genomic information. . Database (Oxford) 2010:, baq013.[PubMed].[CrossRef]
    [Google Scholar]
  8. Cheng Q. , Paszkiet B. J. , Shoemaker N. B. , Gardner J. F. , Salyers A. A. . ( 2000; ). Integration and excision of a Bacteroides conjugative transposon, CTnDOT. . J Bacteriol 182:, 4035–4043. [CrossRef].[PubMed].
    [Google Scholar]
  9. Chiu B. . ( 1999; ). Multiple infections in carotid atherosclerotic plaques. . Am Heart J 138:, S534–S536. [CrossRef].[PubMed].
    [Google Scholar]
  10. Clewell D. B. , Flannagan S. E. . ( 1993; ). The conjugative transposons of Gram-positive bacteria. . In Bacterial Conjugation, pp. 369–393. Edited by Clewell D. B. . . New York:: Plenum Press;.
    [Google Scholar]
  11. Cooper A. J. , Kalinowski A. P. , Shoemaker N. B. , Salyers A. A. . ( 1997; ). Construction and characterization of a Bacteroides thetaiotaomicron recA mutant: transfer of Bacteroides integrated conjugative elements is RecA independent. . J Bacteriol 179:, 6221–6227.[PubMed].
    [Google Scholar]
  12. Jeters R. T. , Wang G. R. , Moon K. , Shoemaker N. B. , Salyers A. A. . ( 2009; ). Tetracycline-associated transcriptional regulation of transfer genes of the Bacteroides conjugative transposon CTnDOT. . J Bacteriol 191:, 6374–6382. [CrossRef].[PubMed].
    [Google Scholar]
  13. Joshipura K. J. , Rimm E. B. , Douglass C. W. , Trichopoulos D. , Ascherio A. , Willett W. C. . ( 1996; ). Poor oral health and coronary heart disease. . J Dent Res 75:, 1631–1636. [CrossRef].[PubMed].
    [Google Scholar]
  14. Kikuchi Y. , Ohara N. , Sato K. , Yoshimura M. , Yukitake H. , Sakai E. , Shoji M. , Naito M. , Nakayama K. . ( 2005; ). Novel stationary-phase-upregulated protein of Porphyromonas gingivalis influences production of superoxide dismutase, thiol peroxidase and thioredoxin. . Microbiology 151:, 841–853. [CrossRef].[PubMed].
    [Google Scholar]
  15. Kondo Y. , Ohara N. , Sato K. , Yoshimura M. , Yukitake H. , Naito M. , Fujiwara T. , Nakayama K. . ( 2010; ). Tetratricopeptide repeat protein-associated proteins contribute to the virulence of Porphyromonas gingivalis . . Infect Immun 78:, 2846–2856. [CrossRef].[PubMed].
    [Google Scholar]
  16. Kozarov E. V. , Dorn B. R. , Shelburne C. E. , Dunn W. A. Jr , Progulske-Fox A. . ( 2005; ). Human atherosclerotic plaque contains viable invasive Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis . . Arterioscler Thromb Vasc Biol 25:, e17–e18. [CrossRef].[PubMed].
    [Google Scholar]
  17. Mattila K. J. , Valtonen V. V. , Nieminen M. , Huttunen J. K. . ( 1995; ). Dental infection and the risk of new coronary events: prospective study of patients with documented coronary artery disease. . Clin Infect Dis 20:, 588–592. [CrossRef].[PubMed].
    [Google Scholar]
  18. Morrison H. I. , Ellison L. F. , Taylor G. W. . ( 1999; ). Periodontal disease and risk of fatal coronary heart and cerebrovascular diseases. . J Cardiovasc Risk 6:, 7–11.[PubMed].
    [Google Scholar]
  19. Naito M. , Hirakawa H. , Yamashita A. , Ohara N. , Shoji M. , Yukitake H. , Nakayama K. , Toh H. , Yoshimura F. et al. ( 2008; ). Determination of the genome sequence of Porphyromonas gingivalis strain ATCC 33277 and genomic comparison with strain W83 revealed extensive genome rearrangements in P. gingivalis. . DNA Res 15:, 215–225. [CrossRef].[PubMed].
    [Google Scholar]
  20. Nakayama K. , Kadowaki T. , Okamoto K. , Yamamoto K. . ( 1995; ). Construction and characterization of arginine-specific cysteine proteinase (Arg-gingipain)-deficient mutants of Porphyromonas gingivalis. Evidence for significant contribution of Arg-gingipain to virulence. . J Biol Chem 270:, 23619–23626.[PubMed].[CrossRef]
    [Google Scholar]
  21. Nelson K. E. , Fleischmann R. D. , DeBoy R. T. , Paulsen I. T. , Fouts D. E. , Eisen J. A. , Daugherty S. C. , Dodson R. J. , Durkin A. S. et al. ( 2003; ). Complete genome sequence of the oral pathogenic bacterium Porphyromonas gingivalis strain W83. . J Bacteriol 185:, 5591–5601. [CrossRef].[PubMed].
    [Google Scholar]
  22. Salyers A. A. , Shoemaker N. B. , Stevens A. M. , Li L. Y. . ( 1995; ). Conjugative transposons: an unusual and diverse set of integrated gene transfer elements. . Microbiol Rev 59:, 579–590.[PubMed].
    [Google Scholar]
  23. Shoemaker N. B. , Getty C. , Guthrie E. P. , Salyers A. A. . ( 1986; ). Regions in Bacteroides plasmids pBFTM10 and pB8-51 that allow Escherichia coliBacteroides shuttle vectors to be mobilized by IncP plasmids and by a conjugative Bacteroides tetracycline resistance element. . J Bacteriol 166:, 959–965.[PubMed].
    [Google Scholar]
  24. Shoemaker N. B. , Wang G. R. , Stevens A. M. , Salyers A. A. . ( 1993; ). Excision, transfer, and integration of NBU1, a mobilizable site-selective insertion element. . J Bacteriol 175:, 6578–6587.[PubMed].
    [Google Scholar]
  25. Song B. , Shoemaker N. B. , Gardner J. F. , Salyers A. A. . ( 2007; ). Integration site selection by the Bacteroides conjugative transposon CTnBST. . J Bacteriol 189:, 6594–6601. [CrossRef].[PubMed].
    [Google Scholar]
  26. Sugawara H. , Ohyama A. , Mori H. , Kurokawa K. . ( 2009; ). Microbial Genome Annotation Pipeline (MiGAP) for diverse users. . In The 20th International Conference on Genome Informatics (GIW2009) , pp. S001-001–S001-002. Yokohama, Japan:: Japanese Society for Bioinformatics;.
    [Google Scholar]
  27. Sutanto Y. , Shoemaker N. B. , Gardner J. F. , Salyers A. A. . ( 2002; ). Characterization of Exc, a novel protein required for the excision of Bacteroides conjugative transposon. . Mol Microbiol 46:, 1239–1246. [CrossRef].[PubMed].
    [Google Scholar]
  28. Sutanto Y. , DiChiara J. M. , Shoemaker N. B. , Gardner J. F. , Salyers A. A. . ( 2004; ). Factors required in vitro for excision of the Bacteroides conjugative transposon, CTnDOT. . Plasmid 52:, 119–130. [CrossRef].[PubMed].
    [Google Scholar]
  29. Taniguchi A. , Nishimura F. , Murayama Y. , Nagasaka S. , Fukushima M. , Sakai M. , Yoshii S. , Kuroe A. , Suzuki H. et al. ( 2003; ). Porphyromonas gingivalis infection is associated with carotid atherosclerosis in non-obese Japanese type 2 diabetic patients. . Metabolism 52:, 142–145. [CrossRef].[PubMed].
    [Google Scholar]
  30. Taylor G. W. . ( 2001; ). Bidirectional interrelationships between diabetes and periodontal diseases: an epidemiologic perspective. . Ann Periodontol 6:, 99–112. [CrossRef].[PubMed].
    [Google Scholar]
  31. Tribble G. D. , Lamont G. J. , Progulske-Fox A. , Lamont R. J. . ( 2007; ). Conjugal transfer of chromosomal DNA contributes to genetic variation in the oral pathogen Porphyromonas gingivalis . . J Bacteriol 189:, 6382–6388. [CrossRef].[PubMed].
    [Google Scholar]
  32. Ueshima J. , Shoji M. , Ratnayake D. B. , Abe K. , Yoshida S. , Yamamoto K. , Nakayama K. . ( 2003; ). Purification, gene cloning, gene expression, and mutants of Dps from the obligate anaerobe Porphyromonas gingivalis . . Infect Immun 71:, 1170–1178. [CrossRef].[PubMed].
    [Google Scholar]
  33. Valentine P. J. , Shoemaker N. B. , Salyers A. A. . ( 1988; ). Mobilization of Bacteroides plasmids by Bacteroides conjugal elements. . J Bacteriol 170:, 1319–1324.[PubMed].
    [Google Scholar]
  34. Waldor M. K. , Tschäpe H. , Mekalanos J. J. . ( 1996; ). A new type of conjugative transposon encodes resistance to sulfamethoxazole, trimethoprim, and streptomycin in Vibrio cholerae O139. . J Bacteriol 178:, 4157–4165.[PubMed].
    [Google Scholar]
  35. Whittle G. , Shoemaker N. B. , Salyers A. A. . ( 2002; a). The role of Bacteroides conjugative transposons in the dissemination of antibiotic resistance genes. . Cell Mol Life Sci 59:, 2044–2054. [CrossRef].[PubMed].
    [Google Scholar]
  36. Whittle G. , Shoemaker N. B. , Salyers A. A. . ( 2002; b). Characterization of genes involved in modulation of conjugal transfer of the Bacteroides conjugative transposon CTnDOT. . J Bacteriol 184:, 3839–3847. [CrossRef].[PubMed].
    [Google Scholar]
  37. Whittle G. , Hamburger N. , Shoemaker N. B. , Salyers A. A. . ( 2006; ). A Bacteroides conjugative transposon, CTnERL, can transfer a portion of itself by conjugation without excising from the chromosome. . J Bacteriol 188:, 1169–1174. [CrossRef].[PubMed].
    [Google Scholar]
  38. Wood M. M. , Dichiara J. M. , Yoneji S. , Gardner J. F. . ( 2010; ). CTnDOT integrase interactions with attachment site DNA and control of directionality of the recombination reaction. . J Bacteriol 192:, 3934–3943. [CrossRef].[PubMed].
    [Google Scholar]
  39. Wozniak R. A. , Waldor M. K. . ( 2010; ). Integrative and conjugative elements: mosaic mobile genetic elements enabling dynamic lateral gene flow. . Nat Rev Microbiol 8:, 552–563. [CrossRef].[PubMed].
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.047803-0
Loading
/content/journal/micro/10.1099/mic.0.047803-0
Loading

Data & Media loading...

Supplements

vol. , part 7, pp. 2022 - 2032

Colonies of strains ATCC 33277 and W83 on eTS agar plates. (Also available in the PDF below.)

Primers used in this study. Transfer frequencies under various conditions. Both tables and a copy of the figure are available as a single PDF(109 KB).



IMAGE
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error