Abstract

A candidate gene analysis of the microphthalmia-associated transcription factor (MITF) gene was used in an attempt to identify the genetic basis for a white-spotted coat color phenotype in the Asian swamp buffalo (Bubalus bubalis carabanensis). Ninety-three buffaloes-32 solid, 38 spotted and 23 white individuals-were Sanger-sequenced for all MITF exons as well as highly conserved intronic and flanking regions. MITF cDNA representing skin and iris tissue from six spotted, nine solid and one white buffaloes was also Sanger-sequenced to confirm detected mutations. Two independent loss-of-function mutations, a premature stop codon (c.328C>T, p.Arg110*) and a donor splice-site mutation (c.840+2T>A, p.Glu281_ Leu282Ins8), both of which cause white-spotted coat color in swamp buffaloes, were identified. The nonsense mutation leads to a premature stop codon in exon 3, and likely removal of the resulting mRNA via nonsense-mediated decay pathway, whereas the donor splice-site mutation leads to aberrant splicing of exon 8 that encodes part of a highly conserved region of MITF. The resulting insertion of eight amino acid residues is expected to perturb the leucine zipper part in the basic helix-loop-helix leucine zipper (bHLH-Zip) domain and will most likely influence dimerization and DNA binding capacity. Electrophoretic mobility shift assay was performed using mutant and wild-type MITF proteins and showed that the mutant MITF protein resulting from the splice-site mutation decreased in vitro DNA binding capacity compared to wild-type MITF. White-spotted buffalo bulls are sacrificed in funeral ceremonies in Tana Toraja, Indonesia, because they are considered holy, and our results show that genetic variation causes a tie to the cultural use of these buffaloes.

Keywords

Bubalus bubalis; donor splice-site mutation; MITF; nonsense mutation

Published in

Animal Genetics
2015, Volume: 46, number: 6, pages: 676-682

SLU Authors

• Yusniza, yulnawati

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    
  • Bogor Agricultural University
  • Indonesian Institute of Sciences (LIPI)

• Wilbe, Maria

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    
  • Uppsala University

• Andersson, Leif

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    
  • Uppsala University

• Andersson, Göran

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Genetics and Breeding


Publication identifier

DOI: https://doi.org/10.1111/age.12334

Permanent link to this page (URI)

https://res.slu.se/id/publ/69257