Sequencing and Characterisation of Rearrangements in Three S. pastorianus Strains Reveals the Presence of Chimeric Genes and Gives Evidence of Breakpoint Reuse

Gross chromosomal rearrangements have the potential to be evolutionarily advantageous to an adapting organism. The generation of a hybrid species increases opportunity for recombination by bringing together two homologous genomes. We sought to define the location of genomic rearrangements in three strains of Saccharomyces pastorianus, a natural lager-brewing yeast hybrid of Saccharomyces cerevisiae and Saccharomyces eubayanus, using whole genome shotgun sequencing. Each strain of S. pastorianus has lost species-specific portions of its genome and has undergone extensive recombination, producing chimeric chromosomes. We predicted 30 breakpoints that we confirmed at the single nucleotide level by designing species-specific primers that flank each breakpoint, and then sequencing the PCR product. These rearrangements are the result of recombination between areas of homology between the two subgenomes, rather than repetitive elements such as transposons or tRNAs. Interestingly, 28/30 S. cerevisiae- S. eubayanus recombination breakpoints are located within genic regions, generating chimeric genes. Furthermore we show evidence for the reuse of two breakpoints, located in HSP82 and KEM1, in strains of proposed independent origin.

[1]  Y. Yoshida,et al.  SSU1-R, a sulfite resistance gene of wine yeast, is an allele of SSU1 with a different upstream sequence , 1998 .

[2]  C. Lopes,et al.  Natural hybrids of S. cerevisiae x S. kudriavzevii share alleles with European wild populations of Saccharomyces kudriavzevii. , 2010, FEMS yeast research.

[3]  G. Liti,et al.  Inferences of evolutionary relationships from a population survey of LTR‐retrotransposons and telomeric‐associated sequences in the Saccharomyces sensu stricto complex , 2005, Yeast.

[4]  M. Aigle,et al.  Molecular genetic study of introgression between Saccharomyces bayanus and S. cerevisiae , 2005, Yeast.

[5]  P. Pevzner,et al.  Dynamics of Mammalian Chromosome Evolution Inferred from Multispecies Comparative Maps , 2005, Science.

[6]  Amparo Querol,et al.  Fermentative stress adaptation of hybrids within the Saccharomyces sensu stricto complex. , 2008, International journal of food microbiology.

[7]  Loretta Auvil,et al.  Breakpoint regions and homologous synteny blocks in chromosomes have different evolutionary histories. , 2009, Genome research.

[8]  D. Dubourdieu,et al.  New Hybrids between Saccharomyces Sensu Stricto Yeast Species Found among Wine and Cider Production Strains , 1998, Applied and Environmental Microbiology.

[9]  G. Sherlock,et al.  Recurrent Rearrangement during Adaptive Evolution in an Interspecific Yeast Hybrid Suggests a Model for Rapid Introgression , 2013, PLoS genetics.

[10]  J. Pérez-Ortín,et al.  Chimeric Genomes of Natural Hybrids of Saccharomyces cerevisiae and Saccharomyces kudriavzevii , 2009, Applied and Environmental Microbiology.

[11]  T. James,et al.  Aneuploidy and copy number breakpoints in the genome of lager yeasts mapped by microarray hybridisation , 2004, Current Genetics.

[12]  M. Ashburner,et al.  Fragile regions and not functional constraints predominate in shaping gene organization in the genus Drosophila. , 2010, Genome research.

[13]  Masahira Hattori,et al.  Genome Sequence of the Lager Brewing Yeast, an Interspecies Hybrid , 2009, DNA research : an international journal for rapid publication of reports on genes and genomes.

[14]  Mark Johnston,et al.  Microbe domestication and the identification of the wild genetic stock of lager-brewing yeast , 2011, Proceedings of the National Academy of Sciences.

[15]  U. Bond Chapter 6: The genomes of lager yeasts. , 2009, Advances in applied microbiology.

[16]  J. Piškur,et al.  A natural chimeric yeast containing genetic material from three species. , 1999, International journal of systematic bacteriology.

[17]  G. Sherlock,et al.  Reconstruction of the genome origins and evolution of the hybrid lager yeast Saccharomyces pastorianus. , 2008, Genome research.

[18]  B. Birren,et al.  Sequencing and comparison of yeast species to identify genes and regulatory elements , 2003, Nature.

[19]  Y. Kaneko,et al.  Pure and Mixed Genetic Lines of Saccharomyces bayanus and Saccharomyces pastorianus and Their Contribution to the Lager Brewing Strain Genome , 2006, Applied and Environmental Microbiology.

[20]  Jose Castresana,et al.  Is mammalian chromosomal evolution driven by regions of genome fragility? , 2006, Genome Biology.

[21]  L. Fulton,et al.  Finding Functional Features in Saccharomyces Genomes by Phylogenetic Footprinting , 2003, Science.

[22]  Andrew W. Murray,et al.  Rapid Expansion and Functional Divergence of Subtelomeric Gene Families in Yeasts , 2010, Current Biology.

[23]  Bernard B. Suh,et al.  Reconstructing contiguous regions of an ancestral genome. , 2006, Genome research.

[24]  J. Nadeau,et al.  Lengths of chromosomal segments conserved since divergence of man and mouse. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[25]  A. Pulvirenti,et al.  The inheritance of mtDNA in lager brewing strains. , 2008, FEMS yeast research.

[26]  P. Barré,et al.  Multiple Ty-mediated chromosomal translocations lead to karyotype changes in a wine strain of Saccharomyces cerevisiae , 1999, Molecular and General Genetics MGG.

[27]  Jack W. Szostak,et al.  The double-strand-break repair model for recombination , 1983, Cell.

[28]  Jun-Yi Leu,et al.  Dynamic Large-Scale Chromosomal Rearrangements Fuel Rapid Adaptation in Yeast Populations , 2013, PLoS genetics.

[29]  P. Pevzner,et al.  Human and mouse genomic sequences reveal extensive breakpoint reuse in mammalian evolution , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[30]  O. Ozier-Kalogeropoulos,et al.  A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae. , 1993, Nucleic acids research.

[31]  P. Brown,et al.  Global mapping of meiotic recombination hotspots and coldspots in the yeast Saccharomyces cerevisiae. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Maitreya J. Dunham,et al.  Competitive Repair by Naturally Dispersed Repetitive DNA during Non-Allelic Homologous Recombination , 2010, PLoS genetics.

[33]  Ruojing Yang,et al.  Glucose Limitation Induces GCN4Translation by Activation of Gcn2 Protein Kinase , 2000, Molecular and Cellular Biology.

[34]  J. Hansen,et al.  Saccharomyces carlsbergensis contains two functional MET2 alleles similar to homologues from S. cerevisiae and S. monacensis. , 1994, Gene.

[35]  Amparo Querol,et al.  Molecular characterization of a chromosomal rearrangement involved in the adaptive evolution of yeast strains. , 2002, Genome research.

[36]  S. Oliver,et al.  Engineering evolution to study speciation in yeasts , 2003, Nature.

[37]  M. Takashio,et al.  Analysis of an inactivated Lg-FLO1 gene present in bottom-fermenting yeast. , 2002, Journal of bioscience and bioengineering.

[38]  A. Martini,et al.  Three newly delimited species of Saccharomyces sensu stricto , 2004, Antonie van Leeuwenhoek.

[39]  Antonis Rokas,et al.  Inferring ancient divergences requires genes with strong phylogenetic signals , 2013, Nature.

[40]  Laureana Rebordinos,et al.  Genome-wide amplifications caused by chromosomal rearrangements play a major role in the adaptive evolution of natural yeast. , 2003, Genetics.

[41]  S. Oliver,et al.  Chromosomal evolution in Saccharomyces , 2000, Nature.

[42]  N. Shirley,et al.  Evidence for multiple interspecific hybridization in Saccharomyces sensu stricto species. , 2002, FEMS yeast research.

[43]  David Botstein,et al.  Characteristic genome rearrangements in experimental evolution of Saccharomyces cerevisiae , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[44]  U. Bond,et al.  Recombination between Homoeologous Chromosomes of Lager Yeasts Leads to Loss of Function of the Hybrid GPH1 Gene , 2009, Applied and Environmental Microbiology.

[45]  T. James,et al.  Lager yeasts possess dynamic genomes that undergo rearrangements and gene amplification in response to stress , 2008, Current Genetics.

[46]  David Collingwood,et al.  Fragile Genomic Sites Are Associated with Origins of Replication , 2009, Genome biology and evolution.