Characterisation of the melanocortin-1 receptor gene in alpaca and identification of possible markers associated with phenotypic variations in colour
N. L. Feeley A and K. A. Munyard A BA School of Biomedical Sciences, Curtin University of Technology, Perth, WA 6845, Australia.
B Corresponding author. Email: k.munyard@exchange.curtin.edu.au
Animal Production Science 49(8) 675-681 https://doi.org/10.1071/AN09005
Submitted: 12 January 2009 Accepted: 28 April 2009 Published: 27 July 2009
Abstract
The aim of this study was to determine if any correlation exists between melanocortin-1 receptor (MC1R) polymorphisms and skin and fibre colour in alpacas. Primers capable of amplifying the entire alpaca MC1R gene were designed from a comparative alignment of Bos taurus and Mus musculus MC1R gene sequences. The complete MC1R gene of 41 alpacas exhibiting a range of fibre colours, and which were sourced from farms across Australia, was sequenced from PCR products. Twenty-one single nucleotide polymorphisms were identified within MC1R. Two of these polymorphisms (A82G and C901T) have the potential to reduce eumelanin production by disrupting the activity of MC1R. No agreement was observed between fibre colour alone and MC1R genotype in the 41 animals in this study. However, when the animals were assigned to groups based on the presence or absence of eumelanin in their fibre and skin, only animals that had at least one allele with the A82/C901 combination expressed eumelanin. We propose that A82/C901 is the wild-type dominant ‘E’ MC1R allele, while alpacas with either G82/T901 or G82/Y901 are homozygous for the recessive ‘e’ MC1R allele and are therefore unable to produce eumelanin.
Additional keywords: melanin, pigmentation, single nucleotide polymorphism.
Acknowledgements
The authors thank the following alpaca breeders for supplying samples from their alpacas, without them this research would not have been possible: Sue Leitch (Brookwood Alpacas), Carolyn Emery (Windella Alpacas), Pinjarra Alpacas, Aviana Farms, Bolero Alpacas, Mangowood Farm, and Tularosa Alpacas. Thanks also to the Australian Alpaca Association for their strong in-kind support of this research. Dr Katherine Morton (Faculty of Veterinary Science, The University of Sydney) kindly provided some of the samples used in this study, and Dr Belinda Appleton (Department of Genetics, The University of Melbourne/Alpaca Genomics Australia), supplied DNA samples for nine of the animals. This research was jointly funded by the Rural Industries Research & Development Corporation and Curtin University. Finally, thanks to Elizabeth Paul (Erehwon Alpacas) for many hours of stimulating colour genetics discussions.
Adan RAH
(2006) Constitutive receptor activity series endogenous inverse agonists and constitutive receptor activity in the melanocortin system. Trends in Pharmacological Sciences 27, 183–186.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Barsh GS
(1996) The genetics of pigmentation: from fancy genes to complex traits. Trends in Genetics 12, 299–305.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Beaumont KA,
Shekar SL,
Newton RA,
James MR,
Stow JL,
Duffy DL, Sturm RA
(2007) Receptor function, dominant negative activity and phenotype correlations for MC1R variant alleles. Human Molecular Genetics 16, 2249–2260.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Busca R, Ballotti R
(2000) Cyclic AMP a key messenger in the regulation of skin pigmentation. Pigment Cell Research 13, 60–69.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Carroll L,
Voisey J, van Daal A
(2005) Gene polymorphisms and their effects in the melanocortin system. Peptides 26, 1871–1885.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Cecchi T,
Cozzali C,
Passamonti P,
Ceccarelli P,
Gargiulo AM,
Frank E, Renieri C
(2004) Melanins and melanosomes from llama. Pigment Cell Research 17, 307–311.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Cecchi T,
Valbonesi A,
Passamonti P,
Frank E, Renieri C
(2007) Quantitative variation of melanins in llama (Lama glama L.). Small Ruminant Research 71, 52–58.
| Crossref | GoogleScholarGoogle Scholar |
Everts RE,
Rothuizen J, AvanOost B
(2000) Identification of a premature stop codon in the melanocyte-stimulating hormone receptor gene (MC1R) in Labrador and Golden retrievers with yellow coat colour. Animal Genetics 31, 194–199.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Fleet MR,
Foulds RA,
Pourbeik T,
McInnes CB,
Smith DH, Burbidge A
(1995) Pigmentation relationships among young Merino sheep and their processed wool. Australian Journal of Experimental Agriculture 35, 343–351.
| Crossref | GoogleScholarGoogle Scholar |
Frandberg P-A,
Doufexis M,
Kapas S, Chhajlani V
(1998) Human pigmentation phenotype: a point mutation generates non-functional MSH receptor. Biochemical and Biophysical Research Communications 245, 490–492.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Frank EN,
Hick MVH,
Gauna CD,
Lamas HE,
Renieri C, Antonini M
(2006) Phenotypic and genetic description of fibre traits in South American domestic camelids (llamas and alpacas). Small Ruminant Research 61, 113–129.
| Crossref | GoogleScholarGoogle Scholar |
Furumura M,
Sakai C,
Abdel-Malek ZA,
Barsh GS, Hearing VJ
(1996) The interaction of Agouti signal protein and melanocyte stimulating hormone to regulate melanin formation in mammals. Pigment Cell Research 9, 191–203.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Furumura M,
Potter SB,
Toyofuku K,
Matsunaga J,
Muller J, Hearing VJ
(2001) Involvement of ITF2 in the transcriptional regulation of melanogenic genes. The Journal of Biological Chemistry 276, 28 147–28 154.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Garcia-Borron JC,
Sanchez BL, Jimenez-Cervantes C
(2005) Melanocortin-1 receptor structure and functional regulation. Pigment Cell Research 18, 393–410.
|
CAS |
PubMed |
Girardot M,
Guibert S,
Laforet M-P,
Gallard Y,
Larroque H, Oulmouden A
(2006) The insertion of a full-length Bos taurus LINE element is responsible for a transcriptional deregulation of the Normande Agouti gene. Pigment Cell Research 19, 346–355.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Grimes EA,
Noake PJ,
Dixon L, Urquhart A
(2001) Sequence polymorphism in the human melanocortin-1 receptor gene as an indicator of the red hair phenotype. Forensic Science International 122, 124–129.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Guibert S,
Girardot M,
Leveziel H,
Julien R, Oulmouden A
(2004) Pheomelanin coat colour dilution in French cattle breeds is not correlated with the TYR, TYRP1 and DCT transcription levels. Pigment Cell Research 17, 337–345.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Hearing VJ
(2005) Biogenesis of pigment granules: a sensitive way to regulate melanocyte function. Journal of Dermatological Science 37, 3–14.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Hoekstra H
(2006) Genetics, development and evolution of adaptive pigmentation in vertebrates. Heredity 97, 222–234.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Hoekstra H,
Hirschmann RJ,
Bundey RA,
Insel PA, Crossland JP
(2006) A single amino acid mutation contributes to adaptive beach mouse colour pattern. Science 313, 101–104.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Hornyak TJ,
Hayes DJ,
Chiu L-Y, Ziff EB
(2001) Transcription factors in melanocyte development: distinct roles for Pax-3 and Mitf. Mechanisms of Development 101, 47–59.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Ishida Y,
David VA,
Eizirik E,
Schaffer AA,
Neelam BA,
Roelke ME,
Hannah SS,
O’Brien SJ, Menotti-Raymond M
(2006) A homozygous single-base deletion in MLPH causes the dilute coat color phenotype in the domestic cat. Genomics 88, 698–705.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Jackson I
(1997) Homologous pigmentation mutations in human, mouse and other model organisms. Human Molecular Genetics 6, 1613–1624.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Jimenez-Cervantes C,
Germer S,
Gonzalez P,
Sanchez J,
Sanchez CO, Garcia-Borron JC
(2001) Thr40 and Met122 are new partial loss-of-function natural mutations of the human melanocortin-1 receptor. FEBS Letters 508, 44–48.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Kerns JA,
Olivier M,
Lust G, Barsh GS
(2003) Exclusion of melanocortin-1 receptor (MC1R) and Agouti as candidates for dominant black in dogs. The Journal of Heredity 94, 75–79.
|
CAS |
PubMed |
Kijas JMH,
Wales R,
Tornsten A,
Chardon P,
Moller M, Andersson L
(1998) Melanocortin-1 receptor (MC1R) mutations and coat colour in pigs. Genetics 150, 1177–1185.
|
CAS |
PubMed |
Krystek S,
Kimura S, Tebben A
(2006) Modeling and active site refinement for G protein-coupled receptors: application to the β-2 adrenergic receptor. Journal of Computer-Aided Molecular Design 20, 463–470.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Le Pape E,
Wakamatsu K,
Ito S,
Wolber R, Hearing VJ
(2008) Regulation of eumelanin/pheomelanin synthesis and visible pigmentation in melanocytes by ligands of the melanocortin-1 receptor. Pigment Cell & Melanoma Research 21, 477–486.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Lin JY, Fisher DE
(2007) Melanocyte biology and skin pigmentation. Nature 445, 843–850.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Logan DW,
Bryson-Richardson RJ,
Pagan KE,
Taylor MS,
Currie PD, Jackson IJ
(2003) The structure and evolution of the melanocortin and MCH receptors in fish and mammals. Genomics 81, 184–191.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Marklund L,
Johansson Moller M,
Sandberg K, Andersson L
(1996) A missense mutation in the gene for melanocyte-stimulating hormone receptor (MC1R) is associated with the chestnut coat colour in horses. Mammalian Genome 7, 895–899.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
McGregor BA
(2006) Production, attributes and relative value of alpaca fleeces in southern Australia and implications for industry development. Small Ruminant Research 61, 93–111.
| Crossref | GoogleScholarGoogle Scholar |
Moro O,
Ideta R, Ifuku O
(1999) Characterization of the promoter region of the human melanocortin-1 receptor (MC1R) gene. Biochemical and Biophysical Research Communications 262, 452–460.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Murisier F,
Guichard S, Beermann F
(2006) A conserved transcriptional enhancer that specifies TYRP1 expression to melanocytes. Developmental Biology 298, 644–655.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Newton JM,
Wilkie AL,
He L,
Jordan SA,
Metallinos DL,
Holmes NG,
Jackson I, Barsh GS
(2000) Melanocortin-1 receptor variation in the domestic dog. Mammalian Genome 11, 24–30.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Newton RA,
Smit SE,
Barnes CC,
Pedley J,
Parsons PG, Sturm RA
(2005) Activation of the cAMP pathway by variant human MC1R alleles expressed in HEK and in melanoma cells. Peptides 26, 1818–1824.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Oyehaug L,
Plahte E,
Vage DI, Omholt SW
(2002) The regulatory basis of melanogenic switching. Journal of Theoretical Biology 215, 449–468.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Paul E
(1999) Theory of colour inheritance in alpacas. Alpacas Australia 28, 10–17.
Potterf SB,
Furumura M,
Sviderskaya EV,
Santis C,
Bennett DC, Hearing VJ
(1998) Normal tyrosine transport and abnormal tyrosinase routing in pink-eyed dilution melanocytes. Experimental Cell Research 244, 319–326.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Powell AJ,
Moss MJ,
Tegland Tree L,
Roeder BL,
Carleton CL,
Campbell E, Kooyman DL
(2008) Characterization of the effect of melanocortin-1 receptor, a member of the hair color gene locus, in alpaca (Lama pacos) fleece color differentiation. Small Ruminant Research 79, 183–187.
| Crossref | GoogleScholarGoogle Scholar |
Rees JL
(2000) The melanocortin-1 receptor (MC1R): more than just red hair. Pigment Cell Research 13, 135–140.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Rees JL
(2003) Genetics of skin and hair colour. Annual Review of Genetics 37, 67–90.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Rieder S,
Taourit S,
Mariat D,
Langlois B, Guérin G
(2001) Mutations in the agouti (ASIP), the extension (MC1R), and the brown (TYRP1) loci and their association to coat color phenotypes in horses (Equus caballus). Mammalian Genome 12, 450–455.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Rouzaud F, Hearing VJ
(2005) Regulatory elements of the melanocortin-1 receptor. Peptides 26, 1858–1870.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Rouzaud F,
Martin J,
Gallett PF,
Delourme D,
Goulemot-Leger V,
Amigues Y,
Ménissier F,
Levéziel H,
Julien R, Oulmouden A
(2000) A first genotyping assay of French cattle breeds on a new allele of the extension gene encoding the melanocortin-1 receptor. Genetics, Selection, Evolution 32, 511–520.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Sanchez-Mas J,
Sanchez-Laorden BL,
Guillo LA,
Jimenez-Cervantes C, Garcia-Borron JC
(2005) The melanocortin-1 receptor carboxyl terminal pentapeptide is essential for MC1R function and expression on the cell surface. Peptides 26, 1848–1857.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Schioth HB,
Haitina T,
Ling MK,
Ringholm A,
Fredriksson R,
Cerda-Reverter JM, Klovins J
(2005) Evolutionary conservation of the structural, pharmacological, and genomic characteristics of the melanocortin receptor subtypes. Peptides 26, 1886–1900.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Schoneberg T,
Schulz A,
Biebermann H,
Hermsdorf T,
Rompler H, Sangkuhl K
(2004) Mutant G-protein-coupled receptors as a cause of human diseases. Pharmacology & Therapeutics 104, 173–206.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Scott MC,
Wakamatsu K,
Ito S,
Kadekaro AL,
Kobayashi N,
Groden J,
Kavanagh R,
Takakuwa T,
Virador V,
Hearing VJ, Abdel-Malek ZA
(2002) Human melanocortin-1 receptor variants, receptor function and melanocyte response to UV radiation. Journal of Cell Science 115, 2349–2355.
|
CAS |
PubMed |
Slominski A,
Tobin DJ,
Shibahara S, Wortsman J
(2004) Melanin pigmentation in mammalian skin and its hormonal regulation. Physiological Reviews 84, 1155–1228.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Smith AG,
Box NF,
Marks LH,
Chen W,
Smit DJ,
Wyeth JR,
Huttley GA,
Easteal S, Sturm RA
(2001) The human melanocortin-1 receptor locus: analysis of transcription unit, locus polymorphism and haplotype evolution. Gene 281, 81–94.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Sponenberg P
(2001) Some educated guesses on colour genetics of alpacas. Alpaca Registry Journal 4, 118–124.
Stitziel NO,
Tseng YY,
Pervouchine D,
Goddeau D,
Kasif S, Liang J
(2003) Structural location of disease-associated single-nucleotide polymorphisms. Journal of Molecular Biology 327, 1021–1030.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Strader CD,
Fong TM,
Tota MR, Underwood D
(1994) Structure and function of G protein coupled receptors. Annual Review of Biochemistry 63, 101–132.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Sturm RA,
Teasdale RD, Box NF
(2001) Human pigmentation genes: identification, structure and consequences of polymorphic variation. Gene 277, 49–62.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Tao Y-X
(2006) Inactivating mutations of G protein-coupled receptors and diseases: structure–function insights and therapeutic implications. Pharmacology & Therapeutics 111, 949–973.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Thiruvenkadan AK,
Kandasamy N, Panneerselvam S
(2008) Coat colour inheritance in horses. Livestock Science 117, 109–129.
| Crossref | GoogleScholarGoogle Scholar |
Tully G
(2007) Genotype versus phenotype: human pigmentation. Forensic Science International; Genetics 1, 105–110.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Vage DI,
Lu DS,
Klungland H,
Lien S,
Adalsteinsson S, Cone RD
(1997) A non-epistatic interaction of Agouti and extension in the fox, Vulpes vulpes. Nature Genetics 15, 311–315.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Vage DI,
Fuglei E,
Snipstad K,
Beheim J,
Landsem VM, Klungland H
(2005) Two cysteine substitutions in the MC1R generate the blue variant of the arctic fox (Alopex lagopus) and prevent expression of the white winter coat. Peptides 26, 1814–1817.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Voisey J,
Box NF, Van Daal A
(2001) A polymorphism study of the human Agouti gene and its association with MC1R. Pigment Cell Research 14, 264–267.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
