Evidence for Recombination in Mycobacterium tuberculosis

ABSTRACT Due to its mostly isolated living environment, Mycobacterium tuberculosis is generally believed to be highly clonal, and thus recombination between different strains must be rare and is not critical for the survival of the species. To investigate the roles recombination could have possibly played in the evolution of M. tuberculosis, an analysis was conducted on previously determined genotypes of 36 synonymous single nucleotide polymorphisms (SNPs) in 3,320 M. tuberculosis isolates. The results confirmed the predominant clonal structure of the M. tuberculosis population. However, recombination between different strains was also suggested. To further resolve the issue, 175 intergenic SNPs and 234 synonymous SNPs were genotyped in 37 selected representative strains. A clear mosaic polymorphic pattern ahead of the MT0105 locus encoding a PPE (Pro-Pro-Glu) protein was obtained, which is most likely a result of recombination hot spot. Given that PPE proteins are thought to be critical in host-pathogen interactions, we hypothesize that recombination has been influential in the history of M. tuberculosis and possibly a major contributor to the diversity observed ahead of the MT0105 locus.

[1]  J. Bull,et al.  An Empirical Test of Bootstrapping as a Method for Assessing Confidence in Phylogenetic Analysis , 1993 .

[2]  B. Efron,et al.  Bootstrap confidence levels for phylogenetic trees. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Pathogen population dynamics: the age of the strain. , 2001, Trends in microbiology.

[4]  P. Hopewell,et al.  Simultaneous infection with two strains of Mycobacterium tuberculosis identified by restriction fragment length polymorphism analysis. , 1999, The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease.

[5]  J. Hein,et al.  A simulation study of the reliability of recombination detection methods. , 2001, Molecular biology and evolution.

[6]  E. Holmes,et al.  Recombination within natural populations of pathogenic bacteria: short-term empirical estimates and long-term phylogenetic consequences. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Martin Wu,et al.  A Mycobacterium avium PPE gene is associated with the ability of the bacterium to grow in macrophages and virulence in mice , 2004, Cellular microbiology.

[8]  K. Derbyshire,et al.  Conjugal transfer of chromosomal DNA in Mycobacterium smegmatis , 1998, Molecular microbiology.

[9]  T. Whittam,et al.  Restricted structural gene polymorphism in the Mycobacterium tuberculosis complex indicates evolutionarily recent global dissemination. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[10]  S. Hasnain,et al.  PPE Antigen Rv2430c of Mycobacterium tuberculosis Induces a Strong B-Cell Response , 2003, Infection and Immunity.

[11]  A. Zharkikh,et al.  Estimation of confidence in phylogeny: the complete-and-partial bootstrap technique. , 1995, Molecular phylogenetics and evolution.

[12]  F. Allerberger,et al.  Simultaneous Infection with Two Drug-SusceptibleMycobacterium tuberculosis Strains in an Immunocompetent Host , 1999, Journal of Clinical Microbiology.

[13]  Joseph Felsenstein,et al.  Is there something wrong with the bootstrap on phylogenies? A reply to Hillis and Bull , 1993 .

[14]  Deborah Jacobs-Sera,et al.  Exploring the Mycobacteriophage Metaproteome: Phage Genomics as an Educational Platform , 2006, PLoS genetics.

[15]  K. Crandall,et al.  Evaluation of methods for detecting recombination from DNA sequences: Computer simulations , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[16]  A. Oskooi Molecular Evolution and Phylogenetics , 2008 .

[17]  S. Salzberg,et al.  Whole-Genome Comparison of Mycobacterium tuberculosis Clinical and Laboratory Strains , 2002, Journal of bacteriology.

[18]  Edward A Graviss,et al.  Genome-wide analysis of synonymous single nucleotide polymorphisms in Mycobacterium tuberculosis complex organisms: resolution of genetic relationships among closely related microbial strains. , 2002, Genetics.

[19]  T. Sauerbruch,et al.  Mixed-strain infection with a drug-sensitive and multidrug-resistant strain of Mycobacterium tuberculosis , 1995, The Lancet.

[20]  Darren Martin,et al.  RDP: detection of recombination amongst aligned sequences , 2000, Bioinform..

[21]  N. Saitou,et al.  The neighbor-joining method: a new method for reconstructing phylogenetic trees. , 1987, Molecular biology and evolution.

[22]  W. Jacobs,et al.  Origins of Highly Mosaic Mycobacteriophage Genomes , 2003, Cell.

[23]  B. Barrell,et al.  Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence , 1998, Nature.

[24]  N. S. Mohan,et al.  Simultaneous infection with multiple strains of Mycobacterium tuberculosis identified by restriction fragment length polymorphism analysis. , 2004, The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease.

[25]  Simon Easteal,et al.  A program for calculating and displaying compatibility matrices as an aid in determining reticulate evolution in molecular sequences , 1996, Comput. Appl. Biosci..

[26]  B. Efron,et al.  Bootstrap confidence levels for phylogenetic trees. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[27]  John Maynard Smith,et al.  Analyzing the mosaic structure of genes , 1992, Journal of Molecular Evolution.

[28]  Vineet Bafna,et al.  Inference about Recombination from Haplotype Data: Lower Bounds and Recombination Hotspots , 2006, J. Comput. Biol..

[29]  M J Sanderson,et al.  Improved bootstrap confidence limits in large-scale phylogenies, with an example from Neo-Astragalus (Leguminosae). , 2000, Systematic biology.

[30]  B. Spratt,et al.  Bacterial population genetics, evolution and epidemiology. , 1999, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[31]  J. M. Smith,et al.  How clonal are bacteria? , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Sudhir Kumar,et al.  MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment , 2004, Briefings Bioinform..

[33]  J. T. Crawford,et al.  Simultaneous infection with multiple strains of Mycobacterium tuberculosis. , 2001, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[34]  S. Cole,et al.  Comparative and functional genomics of the Mycobacterium tuberculosis complex. , 2002, Microbiology.

[35]  T. Ottenhoff,et al.  PPE Protein (Rv3873) from DNA Segment RD1 of Mycobacterium tuberculosis: Strong Recognition of Both Specific T-Cell Epitopes and Epitopes Conserved within the PPE Family , 2003, Infection and Immunity.

[36]  K. Derbyshire,et al.  Unconventional conjugal DNA transfer in mycobacteria , 2003, Nature Genetics.

[37]  D. Burke,et al.  Identification of breakpoints in intergenotypic recombinants of HIV type 1 by bootscanning. , 1995, AIDS research and human retroviruses.

[38]  Thomas L. Madden,et al.  BLAST: at the core of a powerful and diverse set of sequence analysis tools , 2004, Nucleic Acids Res..

[39]  T. Whittam,et al.  Is Mycobacterium tuberculosis 15,000 years old? , 1994, The Journal of infectious diseases.

[40]  R. Brosch,et al.  Ancient Origin and Gene Mosaicism of the Progenitor of Mycobacterium tuberculosis , 2005, PLoS pathogens.

[41]  J. Musser Molecular population genetic analysis of emerged bacterial pathogens: selected insights. , 1996, Emerging infectious diseases.

[42]  A. Zharkikh,et al.  Statistical properties of bootstrap estimation of phylogenetic variability from nucleotide sequences. I. Four taxa with a molecular clock. , 1992, Molecular biology and evolution.

[43]  R. Shafer,et al.  Exogenous reinfection with multidrug-resistant Mycobacterium tuberculosis in an immunocompetent patient. , 1995, Tubercle and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease.

[44]  Walter M. Fitch,et al.  On the Problem of Discovering the Most Parsimonious Tree , 1977, The American Naturalist.

[45]  D. Posada Evaluation of methods for detecting recombination from DNA sequences: empirical data. , 2002, Molecular biology and evolution.

[46]  R. Hudson,et al.  Statistical properties of the number of recombination events in the history of a sample of DNA sequences. , 1985, Genetics.

[47]  W. Fitch Toward Defining the Course of Evolution: Minimum Change for a Specific Tree Topology , 1971 .

[48]  E. Feil,et al.  Population structure and evolutionary dynamics of pathogenic bacteria , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[49]  B. McDonald,et al.  Pathogen population genetics, evolutionary potential, and durable resistance. , 2002, Annual review of phytopathology.

[50]  J. A. Studier,et al.  A note on the neighbor-joining algorithm of Saitou and Nei. , 1988, Molecular biology and evolution.

[51]  S. Sawyer Statistical tests for detecting gene conversion. , 1989, Molecular biology and evolution.

[52]  R. Griffiths,et al.  Bounds on the minimum number of recombination events in a sample history. , 2003, Genetics.

[53]  J. Musser,et al.  Single-nucleotide polymorphism-based population genetic analysis of Mycobacterium tuberculosis strains from 4 geographic sites. , 2006, The Journal of infectious diseases.

[54]  Michael A. Newton,et al.  Bootstrapping phylogenies: Large deviations and dispersion effects , 1996 .

[55]  C. Locht,et al.  Linkage disequilibrium between minisatellite loci supports clonal evolution of Mycobacterium tuberculosis in a high tuberculosis incidence area , 2003, Molecular microbiology.

[56]  P. Fearnhead,et al.  A coalescent-based method for detecting and estimating recombination from gene sequences. , 2002, Genetics.

[57]  N. Smith,et al.  The population structure of Mycobacterium bovis in Great Britain: Clonal expansion , 2003, Proceedings of the National Academy of Sciences of the United States of America.