Evolution of MHC class II SLA‐DRB1 locus in the Croatian wild boar (Sus scrofa) implies duplication and weak signals of positive selection

The wild boar is an ancestor of the domestic pig and an important game species with the widest geographical range of all ungulates. Although a large amount of data are available on major histocompatibility complex (MHC) variability in domestic pigs, only a few studies have been performed on wild boars. Due to their crucial role in appropriate immune responses and extreme polymorphism, MHC genes represent some of the best candidates for studying the processes of adaptive evolution. Here, we present the results on the variability and evolution of the entire MHC class II SLA-DRB1 locus exon 2 in 133 wild boars from Croatia. Using direct sequencing and cloning methods, we identified 20 SLA-DRB1 alleles, including eight new variants, with notable divergence. In some individuals, we documented functional locus duplication, and SLA-DRB1*04:10 was identified as the allele involved in the duplication. The expression of a duplicated locus was confirmed by cloning and sequencing cDNA-derived amplicons. Based on individual genotypes, we were able to assume that alleles SLA-DRB1*04:10 and SLA-DRB1*06:07 are linked as an allelic combination that co-evolves as a two-locus haplotype. Our investigation of evolutionary processes at the SLA-DRB1 locus confirmed the role of intralocus recombination in generating allelic variability, whereas tests of positive selection based on the dN/dS (non-synonymous/synonymous substitution rate ratio) test revealed atypically weak and ambiguous signals.

[1]  V. Cubric-Curik,et al.  Are the dinaric mountains a boundary between continental and mediterranean wild boar populations in Croatia? , 2016, European Journal of Wildlife Research.

[2]  Chankyu Park,et al.  Development of a simultaneous high resolution typing method for three SLA class II genes, SLA-DQA, SLA-DQB1, and SLA-DRB1 and the analysis of SLA class II haplotypes. , 2015, Gene.

[3]  Carlos Fonseca,et al.  Wild boar populations up, numbers of hunters down? A review of trends and implications for Europe. , 2015, Pest management science.

[4]  M. Tector,et al.  Characterization of swine leucocyte antigen alleles in a crossbred pig to be used in xenotransplant studies. , 2014, Tissue antigens.

[5]  Ben Murrell,et al.  IDEPI: Rapid Prediction of HIV-1 Antibody Epitopes and Other Phenotypic Features from Sequence Data Using a Flexible Machine Learning Platform , 2014, PLoS Comput. Biol..

[6]  A. Galov,et al.  Major histocompatibility complex class II variation in bottlenose dolphin from Adriatic Sea: inferences about the extent of balancing selection , 2014 .

[7]  Jiasen Liu,et al.  Characterization of swine leukocyte antigen (SLA) polymorphism by sequence-based and PCR-SSP methods in Chinese Bama miniature pigs. , 2014, Developmental and comparative immunology.

[8]  Koichiro Tamura,et al.  MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. , 2013, Molecular biology and evolution.

[9]  A. Galov,et al.  Extensive polymorphism of the major histocompatibility complex DRA gene in Balkan donkeys: perspectives on selection and genealogy. , 2013, Animal genetics.

[10]  C. Stamatis,et al.  Domestication does not narrow MHC diversity in Sus scrofa , 2013, Immunogenetics.

[11]  Y. Yasukochi,et al.  MHC class II DQB diversity in the Japanese black bear, Ursus thibetanus japonicus , 2012, BMC Evolutionary Biology.

[12]  P. Taberlet,et al.  Evolution of major histocompatibility complex class I and class II genes in the brown bear , 2012, BMC Evolutionary Biology.

[13]  D. Macdonald,et al.  Evolution of MHC class I genes in the European badger (Meles meles) , 2012, Ecology and evolution.

[14]  D. Dechmann,et al.  Independent evolution of functional MHC class II DRB genes in New World bat species , 2012, Immunogenetics.

[15]  K. Seo,et al.  Systematic analysis of swine leukocyte antigen-DRB1 nucleotide polymorphisms using genomic DNA-based high-resolution genotyping and identification of new alleles. , 2011, Tissue antigens.

[16]  M. Apollonio,et al.  Genetic diversity in the European wild boar Sus scrofa: phylogeography, population structure and wild x domestic hybridization , 2011 .

[17]  Vincent Moulton,et al.  RDP3: a flexible and fast computer program for analyzing recombination , 2010, Bioinform..

[18]  W. Babik Methods for MHC genotyping in non‐model vertebrates , 2010, Molecular ecology resources.

[19]  J. Lunney,et al.  Molecular characterization of swine leucocyte antigen class II genes in outbred pig populations. , 2009, Animal genetics.

[20]  G. Bertorelle,et al.  Duplication polymorphism at MHC class II DRB1 locus in the wild boar (Sus scrofa) , 2009, Immunogenetics.

[21]  L. Schook,et al.  Diversification of porcine MHC class II genes: evidence for selective advantage , 2009, Immunogenetics.

[22]  M. Kim,et al.  Sequence‐based characterization of the eight SLA loci in Korean native pigs , 2008, International journal of immunogenetics.

[23]  Ziheng Yang PAML 4: phylogenetic analysis by maximum likelihood. , 2007, Molecular biology and evolution.

[24]  S. Goodman,et al.  Selection and recombination drive the evolution of MHC class II DRB diversity in ungulates , 2006, Heredity.

[25]  J K Lunney,et al.  Nomenclature for factors of the swine leukocyte antigen class II system, 2005. , 2005, Tissue antigens.

[26]  Sergei L. Kosakovsky Pond,et al.  Not so different after all: a comparison of methods for detecting amino acid sites under selection. , 2005, Molecular biology and evolution.

[27]  E. Matisoo-Smith,et al.  Worldwide Phylogeography of Wild Boar Reveals Multiple Centers of Pig Domestication , 2005, Science.

[28]  Sergei L. Kosakovsky Pond,et al.  Datamonkey: rapid detection of selective pressure on individual sites of codon alignments , 2005, Bioinform..

[29]  K. Crandall,et al.  A modified bootscan algorithm for automated identification of recombinant sequences and recombination breakpoints. , 2005, AIDS research and human retroviruses.

[30]  D. Milan,et al.  A physical map of large segments of pig Chromosome 7q11–q14: comparative analysis with human Chromosome 6p21 , 2004, Mammalian Genome.

[31]  S. Sommer,et al.  Evolution of MHC-DRB class II polymorphism in the genus Apodemus and a comparison of DRB sequences within the family Muridae (Mammalia: Rodentia) , 2004, Immunogenetics.

[32]  A. Hughes,et al.  Recent Mammalian Gene Duplications: Robust Search for Functionally Divergent Gene Pairs , 2004, Journal of Molecular Evolution.

[33]  P. Hedrick,et al.  PERSPECTIVE: DETECTING ADAPTIVE MOLECULAR POLYMORPHISM: LESSONS FROM THE MHC , 2003, Evolution; international journal of organic evolution.

[34]  C. Landry,et al.  MHC studies in nonmodel vertebrates: what have we learned about natural selection in 15 years? , 2003, Journal of evolutionary biology.

[35]  Ron Shamir,et al.  The Degenerate Primer Design Problem , 2002, ISMB.

[36]  Mark J. Gibbs,et al.  Sister-Scanning: a Monte Carlo procedure for assessing signals in recombinant sequences , 2000, Bioinform..

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

[38]  M. Nei,et al.  Origins and divergence times of mammalian class II MHC gene clusters. , 2000, The Journal of heredity.

[39]  S. Sawyer,et al.  Possible emergence of new geminiviruses by frequent recombination. , 1999, Virology.

[40]  N. Yuhki,et al.  Comparative genome organization of the major histocompatibility complex: lessons from the Felidae , 1999, Immunological reviews.

[41]  T. Ohta Gene conversion vs point mutation in generating variability at the antigen recognition site of major histocompatibility complex loci , 1995, Journal of Molecular Evolution.

[42]  D. Wiley,et al.  Three-dimensional structure of the human class II histocompatibility antigen HLA-DR1 , 1993, Nature.

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

[44]  M. Nei,et al.  Simple methods for estimating the numbers of synonymous and nonsynonymous nucleotide substitutions. , 1986, Molecular biology and evolution.

[45]  J. Kusak,et al.  Extensive polymorphism and evidence of selection pressure on major histocompatibility complex DLA-DRB1, DQA1 and DQB1 class II genes in Croatian grey wolves. , 2013, Tissue antigens.

[46]  J. Trowsdale,et al.  Comparative genomics of major histocompatibility complexes , 2004, Immunogenetics.

[47]  T. A. Hall,et al.  BIOEDIT: A USER-FRIENDLY BIOLOGICAL SEQUENCE ALIGNMENT EDITOR AND ANALYSIS PROGRAM FOR WINDOWS 95/98/ NT , 1999 .

[48]  E. Randi,et al.  The systematics of the wild boar (Sus scrofa L.) in Italy , 1988 .

[49]  T. Jukes CHAPTER 24 – Evolution of Protein Molecules , 1969 .

[50]  Sergei L. Kosakovsky Pond,et al.  BIOINFORMATICS APPLICATIONS , 2022 .

[51]  J. Traherne,et al.  Review Article doi: 10.1111/j.1744-313X.2008.00765.x Blackwell , 2022 .

[52]  S. Frost,et al.  Bioinformatics Applications Note Sequence Analysis Datamonkey 2010: a Suite of Phylogenetic Analysis Tools for Evolutionary Biology , 2022 .