This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Stinear, T. P. Articles by Cole, S. T. Search for Related Content PubMed PubMed Citation Articles by Stinear, T. P. Articles by Cole, S. T.

 Previous Article  |  Next Article 

Journal of Bacteriology, March 2005, p. 1668-1676, Vol. 187, No. 5
0021-9193/05/$08.00+0     doi:10.1128/JB.187.5.1668-1676.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Common Evolutionary Origin for the Unstable Virulence Plasmid pMUM Found in Geographically Diverse Strains of Mycobacterium ulcerans

Timothy P. Stinear,^1,2* Hui Hong,³ Wafa Frigui,¹ Melinda J. Pryor,^4, Roland Brosch,¹ Thierry Garnier,¹ Peter F. Leadlay,⁵ and Stewart T. Cole¹

Unité de Génétique Moléculaire Bactérienne,¹ Plate-Forme 4-Intégration et Analyse Génomiques, Génopole, Institut Pasteur, Paris, France,⁴ Department of Microbiology, Monash University, Clayton, Australia,² Department of Chemistry,³ Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom⁵

Received 8 October 2004/ Accepted 29 November 2004

The 174-kb virulence plasmid pMUM001 in Mycobacterium ulcerans epidemic strain Agy99 harbors three very large and homologous genes that encode giant polyketide synthases (PKS) responsible for the synthesis of the lipid toxin mycolactone. Deeper investigation of M. ulcerans Agy99 resulted in identification of two types of spontaneous deletion variants of pMUM001 within a population of cells that also contained the intact plasmid. These variants arose from recombination between two 8-kb sections of the same plasmid sequence, resulting in the loss of a 65-kb region bearing two of the three mycolactone PKS genes. Investigation of nine diverse M. ulcerans strains by using PCR and Southern hybridization for eight pMUM001 gene sequences confirmed the presence of pMUM001-like elements (collectively called pMUM) in all M. ulcerans strains. Physical mapping of these plasmids revealed that like M. ulcerans Agy99, three strains had undergone major deletions in their mycolactone PKS loci. Online liquid chromatography-sequential mass spectrometry analysis of lipid extracts confirmed that strains with PKS deletions were unable to produce mycolactone or any related cometabolites. Interstrain comparisons of the plasmid gene sequences revealed greater than 98% nucleotide identity, and the phylogeny inferred from these sequences closely mimicked the phylogeny from a previous multilocus sequence typing study in which chromosomally encoded loci were used, a result that is consistent with the hypothesis that M. ulcerans diverged from the closely related organism Mycobacterium marinum by acquiring pMUM. Our results suggest that pMUM is a defining characteristic of M. ulcerans but that in the absence of purifying selection, deletion of plasmid sequences and a corresponding loss of mycolactone production readily arise.

Present address: Department of Biochemistry and Molecular Biology, Monash University, Clayton 3800, Australia.

This article has been cited by other articles:

• Huber, C. A., Ruf, M.-T., Pluschke, G., Kaser, M. (2008). Independent Loss of Immunogenic Proteins in Mycobacterium ulcerans Suggests Immune Evasion. CVI 15: 598-606 [Abstract] [Full Text]  
• Nakanaga, K., Ishii, N., Suzuki, K., Tanigawa, K., Goto, M., Okabe, T., Imada, H., Kodama, A., Iwamoto, T., Takahashi, H., Saito, H. (2007). "Mycobacterium ulcerans subsp. shinshuense" Isolated from a Skin Ulcer Lesion: Identification Based on 16S rRNA Gene Sequencing. J. Clin. Microbiol. 45: 3840-3843 [Abstract] [Full Text]  
• Torrado, E., Adusumilli, S., Fraga, A. G., Small, P. L. C., Castro, A. G., Pedrosa, J. (2007). Mycolactone-Mediated Inhibition of Tumor Necrosis Factor Production by Macrophages Infected with Mycobacterium ulcerans Has Implications for the Control of Infection. Infect. Immun. 75: 3979-3988 [Abstract] [Full Text]  
• Yip, M. J., Porter, J. L., Fyfe, J. A. M., Lavender, C. J., Portaels, F., Rhodes, M., Kator, H., Colorni, A., Jenkin, G. A., Stinear, T. (2007). Evolution of Mycobacterium ulcerans and Other Mycolactone-Producing Mycobacteria from a Common Mycobacterium marinum Progenitor. J. Bacteriol. 189: 2021-2029 [Abstract] [Full Text]  
• Torrado, E., Fraga, A. G., Castro, A. G., Stragier, P., Meyers, W. M., Portaels, F., Silva, M. T., Pedrosa, J. (2007). Evidence for an Intramacrophage Growth Phase of Mycobacterium ulcerans. Infect. Immun. 75: 977-987 [Abstract] [Full Text]  
• Stinear, T. P., Seemann, T., Pidot, S., Frigui, W., Reysset, G., Garnier, T., Meurice, G., Simon, D., Bouchier, C., Ma, L., Tichit, M., Porter, J. L., Ryan, J., Johnson, P. D.R., Davies, J. K., Jenkin, G. A., Small, P. L.C., Jones, L. M., Tekaia, F., Laval, F., Daffe, M., Parkhill, J., Cole, S. T. (2007). Reductive evolution and niche adaptation inferred from the genome of Mycobacterium ulcerans, the causative agent of Buruli ulcer. Genome Res 17: 192-200 [Abstract] [Full Text]  
• Oliveira, M. S., Fraga, A. G., Torrado, E., Castro, A. G., Pereira, J. P., Filho, A. L., Milanezi, F., Schmitt, F. C., Meyers, W. M., Portaels, F., Silva, M. T., Pedrosa, J. (2005). Infection with Mycobacterium ulcerans Induces Persistent Inflammatory Responses in Mice. Infect. Immun. 73: 6299-6310 [Abstract] [Full Text]