Abstract
The gene for agouti signaling protein (ASIP) is centrally involved in the expression of coat color traits in animals. The Mangalitza pig breed is characterized by a black-and-tan phenotype with black dorsal pigmentation and yellow or white ventral pigmentation. We investigated a Mangalitza × Piétrain cross and observed a coat color segregation pattern in the F2 generation that can be explained by virtue of two alleles at the MC1R locus and two alleles at the ASIP locus. Complete linkage of the black-and-tan phenotype to microsatellite alleles at the ASIP locus on SSC 17q21 was observed. Corroborated by the knowledge of similar mouse coat color mutants, it seems therefore conceivable that the black-and-tan pigmentation of Mangalitza pigs is caused by an ASIP allele at, which is recessive to the wild-type allele A. Toward positional cloning of the at mutation, a 200-kb genomic BAC/PAC contig of this chromosomal region has been constructed and subsequently sequenced. Full-length ASIP cDNAs obtained by RACE differed in their 5′ untranslated regions, whereas they shared a common open reading frame. Comparative sequencing of all ASIP exons and ASIP cDNAs between Mangalitza and Piétrain pigs did not reveal any differences associated with the coat color phenotype. Relative qRT-PCR analyses showed different dorsoventral skin expression intensities of the five ASIP transcripts in black-and-tan Mangalitza. The at mutation is therefore probably a regulatory ASIP mutation that alters its dorsoventral expression pattern.
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References
Abecasis GR, Cherny SS, Cookson WO, Cardon LR (2002) Merlin—rapid analysis of dense genetic maps using sparse gene flow trees. Nature Genet 30: 97–101
Al-Bayati H, Duscher S, Kollers S, Rettenberger G, Fries R, et al. (1999) Construction and characterization of a porcine P1-derived artificial chromosome (PAC) library covering 3.2 genome equivalents and cytogenetical assignment of six type I and type II loci. Mamm Genome 10: 569–572
Bultman SJ, Michaud EJ, Woychik RP (1992) Molecular characterization of the mouse agouti locus. Cell 71: 1195–1204
Bultman SJ, Klebig ML, Michaud EJ, Sweet HO, Davisson MT, et al. (1994) Molecular analysis of reverse mutations from nonagouti (a) to black-and-tan (at) and white-bellied agouti (Aw) reveals alternative forms of agouti transcripts. Genes Dev 8: 481–490
Candille SI, Van Raamsdonk CD, Chen C, Kuijper S, Chen-Tsai Y, et al. (2004) Dorsoventral patterning of the mouse coat by Tbx15. PLoS Biol 2: E3
Carroll L, Voisey J, van Daal A (2004) Mouse models of obesity. Clin Dermatol 22: 345–349
Chen Y, Duhl DM, Barsh GS (1996) Opposite orientations of an inverted duplication and allelic variation at the mouse agouti locus. Genetics 144: 265–277
Ellegren H, Chowdhary BP, Johansson M, Marklund L, Fredholm M, et al. (1994) A primary linkage map of the porcine genome reveals a low rate of genetic recombination. Genetics 137: 1089–1100
Girardot M, Martin J, Guibert S, Leveziel H, Julien R, et al. (2005) Widespread expression of the bovine Agouti gene results from at least three alternative promoters. Pigment Cell Res 18: 34–41
Harris RB, Mitchell TD, Mynatt RL (2002) Leptin responsiveness in mice that ectopically express agouti protein. Physiol Behav 75: 159–167
Huszar D, Lynch CA, Fairchild-Huntress V, Dunmore JH, Fang Q, et al. (1997) Targeted disruption of the melanocortin-4 receptor results in obesity in mice. Cell 88: 131–141
Kerns JA, Newton J, Berryere TG, Rubin EM, Cheng JF, et al. (2004) Characterization of the dog Agouti gene and a nonagoutimutation in German Shepherd Dogs. Mamm Genome 15: 798–808
Kijas JMH, Törnsten A, Chowdhary B, Andersson L (1998a) Porcine agouti gene map position SSC 17q21. Chromosome Res 6: 243
Kijas JMH, Wales R, Törnsten A, Chardon P, Moller M, et al. (1998b) Melanocortin receptor 1 (MC1R) mutations and coat color in pigs. Genetics 150: 1177–1185
Kijas JMH, Moller M, Plastow G, Andersson L (2001) A frameshift mutation in MC1R and a high frequency of somatic reversions cause black spotting in pigs. Genetics 158: 779–785
Kwon HY, Bultman SJ, Loffler C, Chen WJ, Furdon PJ, et al. (1994) Molecular structure and chromosomal mapping of the human homolog of the agouti gene. Proc Natl Acad Sci U S A 91: 9760–9764
Leeb T, Rohrer GA (2002) Characterization and chromosome assignment of the porcine AHCY gene for S-adenosylhomocysteine hydrolase. Cytogenet Genome Res 97: 116–119
Leeb T, Deppe A, Kriegesmann B, Brenig B (2000) Genomic structure and nucleotide polymorphisms of the porcine agouti signalling protein gene (ASIP). Anim Genet 31: 335–336
Lu D, Willard D, Patel IR, Kadwell S, Overton L, et al. (1994) Agouti protein is an antagonist of the melanocyte-stimulating-hormone receptor. Nature 371: 799–802
Martins-Wess F, Voß–Nemitz R, Drögemüller C, Brenig B, Leeb T (2002) Construction of a 1.2-Mb BAC/PAC Contig of the porcine gene RYR1 Region on SSC 6q1.2 and comparative analysis with HSA 19q13.13. Genomics 80: 416–422
Millar SE, Miller MW, Stevens ME, Barsh GS (1995) Expression and transgenic studies of the mouse agouti gene provide insight into the mechanisms by which mammalian coat color patterns are generated. Development 121: 3223–3232
Silvers WK, Russel ES (1955) An experimental approach to action of genes at the agouti locus in mouse. J Exp Zool 130: 199–220
23.Siracusa LD (1994) The agouti gene: turned on to yellow. Trends Genet 10, 423–428
Vage DI, Lu D, Klungland H, Lien S, Adalsteinsson S, et al. (1997) A non-epistatic interaction of agouti and extension in the fox, Vulpes vulpes. Nat Genet 15: 311–315
Vrieling H, Duhl DM, Millar SE, Miller KA, Barsh GS (1994) Differences in dorsal and ventral pigmentation result from regional expression of the mouse agouti gene. Proc Natl Acad Sci U S A 91: 5667–5671
Wiedemann S, Kijas J, Andersson L, Fries R (2000) Genetic mapping of agouti in a Mangalitza x Piétrain cross. Proceedings of the 27th International Conference on Animal Genetics, Minnesota, July 22–26, 2000 , B060, 37
Acknowledgments
The authors thank D. Seinige, H. Klippert–Hasberg, S. Neander, and B. Hayn for expert technical assistance, the staff of the Experimental Station Thalhausen of the Technical University of Munich for careful animal husbandry, and D. Fries for the color phenotyping of F2 animals. They also thank P. de Jong and his lab for providing the RPCI-44 library and excellent supplementary information. This study was supported by a grant of the German Research Council DFG (Le 1032/12-1) to TL.
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Nucleotide sequence data reported are available in the EMBL database under accession numbers AJ427478, AJ634673– AJ634677, and AM050718.
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Drögemüller, C., Giese, A., Martins-Wess, F. et al. The mutation causing the black-and-tan pigmentation phenotype of Mangalitza pigs maps to the porcine ASIP locus but does not affect its coding sequence. Mamm Genome 17, 58–66 (2006). https://doi.org/10.1007/s00335-005-0104-1
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DOI: https://doi.org/10.1007/s00335-005-0104-1