title: New insights into the melanophilin (MLPH) gene controlling coat color phenotypes in American mink.
abstract: The mutation causing the Silverblue color type (pp) is one of the most used recessive mutations within American mink (Neovison vison) fur farming, since it is involved in some of the popular color types such as Violet and Saphire which originate from a combination of recessive mutations. In the present study, the genomic and mRNA sequences of the melanophilin (MLPH) gene were studied in Violet, Silverblue and wild-type (wt) mink animals. Although breeding schemes and previous literature indicates that the Violet (aammpp) phenotype is a triple recessive color type involving the same locus as the Silverblue (pp) color type, our findings indicate different genotypes at the MLPH locus. Upon comparison at genomic level, we identified two deletions of the entire intron 7 and of the 5' end of intron 8 in the sequence of the Silverblue MLPH gene. When investigating the mRNA, the Silverblue animals completely lack exon 8, which encodes 65 residues, of which 47 define the Myosin Va (MYO5A) binding domain. This may cause the incorrect anchoring of the MLPH protein to MYO5A in Silverblue animals, resulting in an improper pigmentation as seen in diluted phenotypes. Additionally, in the MLPH mRNA of wt, Violet and Silverblue phenotypes, part of intron 8 is retained resulting in a truncated MLPH protein, which is 359 residues long in wt and Violet and 284 residues long in Silverblue. Subsequently, our findings point out that the missing actin-binding domain, in neither of the 3 analyzed phenotypes affects the transport of melanosomes or the consequent final pigmentation. Moreover, the loss of the major part of the MYO5A domain in the Silverblue MLPH protein seems to be the responsible for the dilute phenotype. Based on our genomic DNA data, genetic tests for selecting Silverblue and Violet carrier animals can be performed in American mink.
authors.affiliation: University of Copenhagen, Faculty of Health and Medical Science, Department of Veterinary Clinical and Animal Sciences, Division of Genetics, Bioinformatics and Breeding, 1870 Frederiksberg C, Denmark.
authors.forename: Susanna
authors.initials: S
authors.lastname: Cirera
authors.forename: Marios N
authors.initials: MN
authors.lastname: Markakis
authors.forename: Knud
authors.initials: K
authors.lastname: Christensen
authors.forename: Razvan
authors.initials: R
authors.lastname: Anistoroaei
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D004252:DNA Mutational Analysis; D005091:Exons; D005808:Genes, Recessive; D056726:Genetic Association Studies; D006197:Hair / Q000502:physiology; D006200:Hair Color / Q000235:genetics*; D018895:Microsatellite Repeats; D008907:Mink / Q000235:genetics; D008969:Molecular Sequence Data; D010375:Pedigree; D010641:Phenotype; D054730:Protein Interaction Domains and Motifs; D012333:RNA, Messenger / Q000235:genetics; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D012333:RNA, Messenger
keywords: American mink; Asparagine; BAC; BLAST; Bacterial artificial chromosome; Basic Local Alignment Search Tool; Bluish Sapphire genotype; Complementary DNA; Deletion; DeltaAB; G; Glutamine; H; Histidine; KA; LOD; LYST; MLPH; MYO5A; Myosin Va; N; NCBI; National Center for Biotechnology Information; Neovison vison; ORF; PCR; Pearl genotype; Polymerase Chain Reaction; RAB27A; S; SNP; Sdw; Serine; Silverblue; Silverblue genotype; Slac2-a deletion mutant; Slac2-a point mutant; Steelblue genotype; UTR; Violet; Violet genotype; aammpp; aapp; base pair; bp; cDNA; logarithm (base 10) of odds; lysosomal trafficking regulator gene; mRNA; melanophilin; member RAS oncogene family; messenger RNA; mg; milligrams; nanograms; ng; nt; nucleotides; open reading frame; p(s)p(s); pp; ppmm; single nucleotide polymorphism; sterile distillated water; untranslated region; wild-type; wt