Mammalian Genome

, Volume 15, Issue 10, pp 798–808 | Cite as

Characterization of the dog Agouti gene and a nonagoutimutation in German Shepherd Dogs

  • Julie A. Kerns
  • J. Newton
  • Tom G. Berryere
  • Edward M. Rubin
  • Jan-Fang Cheng
  • Sheila M. Schmutz
  • Gregory S. BarshEmail author
Original Contributions


The interaction between two genes, Agouti and Melanocortin-1 receptor (Mc1r), produces diverse pigment patterns in mammals by regulating the type, amount, and distribution pattern of the two pigment types found in mammalian hair: eumelanin (brown/black) and pheomelanin (yellow/red). In domestic dogs (Canis familiaris), there is a tremendous variation in coat color patterns between and within breeds; however, previous studies suggest that the molecular genetics of pigment-type switching in dogs may differ from that of other mammals. Here we report the identification and characterization of the Agouti gene from domestic dogs, predicted to encode a 131-amino-acid secreted protein 98% identical to the fox homolog, and which maps to chromosome CFA24 in a region of conserved linkage. Comparative analysis of the Doberman Pinscher Agouti cDNA, the fox cDNA, and 180 kb of Doberman Pinscher genomic DNA suggests that, as with laboratory mice, different pigment-type-switching patterns in the canine family are controlled by alternative usage of different promoters and untranslated first exons. A small survey of Labrador Retrievers, Greyhounds, Australian Shepherds, and German Shepherd Dogs did not uncover any polymorphisms, but we identified a single nucleotide variant in black German Shepherd Dogs predicted to cause an Arg-to-Cys substitution at codon 96, which is likely to account for recessive inheritance of a uniform black coat.


Coat Color Labrador Retriever Golden Retriever Doberman Pinscher Agouti Locus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank J. Faraco and E. Mignot for providing the canine BAC library, and J. Longmire for his support of JAK. We are grateful to the dog breeders who generously submitted DNA samples from their litters and to the DogMap and the FHCRC dog genome project for providing public access to the canine map and marker data at index.html and dog_genome/ .


  1. Barsh, GS 1996The genetics of pigmentation: from fancy genes to complex traitsTrends Genet12 299305CrossRefPubMedGoogle Scholar
  2. Bolin, KA, Anderson, DJ, Trulson, JA, Thompson, DA, Wilken, J,  et al. 1999NMR structure of a minimized human agouti related protein prepared by total chemical synthesisFEBS Lett451125131CrossRefPubMedGoogle Scholar
  3. Breen, M, Thomas, R, Binns, MM, Carter, NP, Langford, CF 1999Reciprocal chromosome painting reveals detailed regions of conserved synteny between the karyotypes of the domestic dog (Canis familiaris) and humanGenomics61145155CrossRefPubMedGoogle Scholar
  4. Bultman, SJ, Michaud, EJ, Woychik, RP 1992Molecular characterization of the mouse agouti locusCell7111951204PubMedGoogle Scholar
  5. Bultman, SJ, Klebig, ML, Michaud, EJ, Sweet, HO, Davisson, MT,  et al. 1994Molecular analysis of reverse mutations from nonagouti (A) to Black-and-Tan (A(T)) and White-Bellied agouti (A(W)) reveals alternative forms of agouti transcriptsGene Dev8481490PubMedGoogle Scholar
  6. Burns, M, Fraser, MN 1966Genetics of the Dog. The basis of successful breedingOliver and BoydEdinburgh, ScotlandGoogle Scholar
  7. Candille, SI, Raamsdonk, CD, Chen, C, Kuijper, S, Chen–Tsai, Y,  et al. 2004Dorsoventral patterning of the mouse coat by Tbx15PLoS Biol23042CrossRefGoogle Scholar
  8. Carver, EA 1984Coat color genetics of the German Shepherd DogJ Hered75247252Google Scholar
  9. Eizirik, RE, Yuhki, N, Johnson, WE, Menotti–Raymond, M, Hannah, SS,  et al. 2003Molecular genetics and evolution of melanism in the cat familyCurr Biol13448453CrossRefPubMedGoogle Scholar
  10. Everts, RE, Rothuizen, J, Oost, BA 2000Identification of a premature stop codon in the melanocyte-stimulating hormone receptor gene (MC1R) in Labrador and Golden retrievers with yellow coat colourAnim Genet31194199CrossRefPubMedGoogle Scholar
  11. Ha, T, Naysmith, L, Waterston, K, Oh, C, Weller, R,  et al. 2003Defining the quantitative contribution of the melanocortin 1 receptor (MC1R) to variation in pigmentary phenotypeAnn NY Acad Sci994339347PubMedGoogle Scholar
  12. Hustad, CM, Perry, WL, Siracusa, LD, Rasberry, C, Cobb, L,  et al. 1995Molecular genetic characterization of six recessive viable alleles of the mouse agouti locusGenetics140255265PubMedGoogle Scholar
  13. Iljin, NA 1941Wolf-Dog GeneticsJ Genet42359427Google Scholar
  14. Jackson, IJ 1994Molecular and developmental genetics of mouse coat colorAnnu Rev of Genet28189217CrossRefGoogle Scholar
  15. Joerg, H, Fries, HR, Meijerink, E, Stranzinger, GF 1996Red coat color in Holstein cattle is associated with a deletion in the MSHR geneMamm Genome7317318CrossRefPubMedGoogle Scholar
  16. Jordan, SA, Jackson, IJ 1998Melanocortin receptors and antagonists regulate pigmentation and body weightBioessays20603606CrossRefPubMedGoogle Scholar
  17. Kerns, JA, Olivier, M, Lust, G, Barsh, GS 2003Exclusion of melanocortin-1 receptor (mc1r) and agouti as candidates for dominant black in dogsJ Hered947579PubMedGoogle Scholar
  18. Kiefer, LL, Ittoop, ORR, Bunce, K, Truesdale, AT, Willard, DH,  et al. 1997Mutations in the carboxyl terminus of the agouti protein decrease agouti inhibition of ligand binding to the melanocortin receptorsBiochemistry3620842090CrossRefPubMedGoogle Scholar
  19. Kijas, JM, Wales, R, Tornsten, A, Chardon, P, Moller, M,  et al. 1998Melanocortin receptor 1 (MC1R) mutations and coat color in pigsGenetics15011771185PubMedGoogle Scholar
  20. Klungland, H, Vage, DI, Gomez–Raya, L, Adalsteinsson, S, Lien, S 1995The role of melanocyte-stimulating hormone (MSH) receptor in bovine coat color determinationMamm Genome6636639CrossRefPubMedGoogle Scholar
  21. Kuramoto, T, Nomoto, T, Sugimura, T, Ushijima, T 2001Cloning of the rat agouti gene and identification of the rat nonagouti mutationMamm Genome12469471CrossRefPubMedGoogle Scholar
  22. Li, R, Mignot, E, Faraco, J, Kadotani, H, Cantanese, J,  et al. 1999Construction and characterization of an eightfold redundant dog genomic bacterial artificial chromosome libraryGenomics58917CrossRefPubMedGoogle Scholar
  23. Ling, MK, Lagerstrom, MC, Fredriksson, R, Okimoto, R, Mundy, NI,  et al. 2003Association of feather colour with constitutively active melanocortin 1 receptors in chickenEur J Biochem27014411449PubMedGoogle Scholar
  24. Lingaas, F, Sorensen, A, Juneja, RK, Johansson, S, Fredholm, M,  et al. 1997Towards construction of a canine linkage map: establishment of 16 linkage groupsMamm Genome8218221CrossRefPubMedGoogle Scholar
  25. Little, CC 1957The inheritance of coat color in dogsComstockIthaca, N.Y.Google Scholar
  26. Lu, DS, Willard, D, Patel, IR, Kadwell, S, Overton, L,  et al. 1994Agouti protein is an antagonist of the melanocyte-stimulating-hormone receptorNature371799802CrossRefPubMedGoogle Scholar
  27. Marklund, L, Moller, MJ, Sandberg, K, Andersson, L 1996A missense mutation in the gene for melanocyte-stimulating hormone receptor (MC1R) is associated with the chestnut coat color in horsesMamm Genome7895899CrossRefPubMedGoogle Scholar
  28. McNulty, JC, Thompson, DA, Bolin, KA, Wilken, J, Barsh, GS,  et al. 2001High-resolution NMR structure of the chemically-synthesized melanocortin receptor binding domain AGRP(87-132) of the agouti-related proteinBiochemistry401552015527CrossRefPubMedGoogle Scholar
  29. Miller, MW, Duhl, DMJ, Vrieling, H, Cordes, SP, Ollmann, MM,  et al. 1993Cloning of the mouse agouti gene predicts a secreted protein ubiquitously expressed in mice carrying the Lethal-Yellow mutationGenes Dev7454467PubMedGoogle Scholar
  30. Miltenberger, RJ, Wakamatsu, K, Ito, S, Woychik, RP, Russell, LB,  et al. 2002Molecular and phenotypic analysis of 25 recessive, homozygous-viable alleles at the mouse agouti locusGenetics160659674PubMedGoogle Scholar
  31. Mundy, NI, Kelly, J, Theron, E, Hawkins, K 2003Evolutionary genetics of the melanocortin-1 receptor in vertebratesAnn NY Acad Sci994307312PubMedGoogle Scholar
  32. Newton, JM, Wilkie, AL, He, L, Jordan, SA, Metallinos, DL,  et al. 2000Melanocortin 1 receptor variation in the domestic dogMamm Genome112430CrossRefPubMedGoogle Scholar
  33. Ollmann, MM, Wilson, BD, Yang, YK, Kerns, JA, Chen, Y,  et al. 1997Antagonism of central melanocortin receptors in vitro and in vivo by agouti-related proteinScience278135138CrossRefPubMedGoogle Scholar
  34. Ollmann, MM, Lamoreux, ML, Wilson, BD, Barsh, GS 1998Interaction of Agouti protein with the melanocortin 1 receptor in vitro and in vivoGenes Dev12316330PubMedGoogle Scholar
  35. Perry, WL, Hustad, CM, Swing, DA, Jenkins, NA, Copeland, NG 1995A transgenic mouse assay for agouti protein activityGenetics140267274PubMedGoogle Scholar
  36. Rieder, S, Taourit, S, Mariat, D, Langlois, B, Guerin, G 2001Mutations in the agouti (ASIP), the extension (MC1R), and the brown (TYRP1) loci and their association to coat color phenotypes in horses (Equus caballus)Mamm Genome12450455CrossRefPubMedGoogle Scholar
  37. Ritland, K, Newton, C, Marshall, HD 2001Inheritance and population structure of the white-phased “Kermode” black bearCurr Biol1114681472CrossRefPubMedGoogle Scholar
  38. Robbins, LS, Nadeau, JH, Johnson, KR, Kelly, MA, Rosellirehfuss, L,  et al. 1993Pigmentation phenotypes of variant extension locus alleles result from point mutations that alter MSH receptor functionCell72 827834CrossRefPubMedGoogle Scholar
  39. Savolainen, P, Zhang, YP, Luo, J, Lundeberg, J, Leitner, T 2002Genetic evidence for an East Asian origin of domestic dogsScience298 16101613CrossRefPubMedGoogle Scholar
  40. Schmutz, SM, Berryere, TG, Ellinwood, NM, Kerns, JA, Barsh, GS 2003MC1R studies in dogs with melanistic mask or brindle patternsJ Hered946973CrossRefPubMedGoogle Scholar
  41. Searle, AG 1968Comparative Genetics of Coat Color in MammalsAcademic PressNew YorkGoogle Scholar
  42. Silvers, WK 1979The Coat Colors of Mice: A Model for Mammalian Gene Action and InteractionSpringer-VerlagNew YorkGoogle Scholar
  43. Sturm, RA, Duffy, DL, Box, NF, Newton, RA, Shepherd, AG,  et al. 2003Genetic association and cellular function of MC1R variant alleles in human pigmentationAnn NY Acad Sci994348358PubMedGoogle Scholar
  44. Takeuchi, S, Suzuki, H, Yabuuchi, M, Takahashi, S 1996A possible involvement of melanocortin-1 receptor in regulating feather color pigmentation in the chickenBiochim Biophys Acta1308164168PubMedGoogle Scholar
  45. Theron, E, Hawkins, K, Bermingham, E, Ricklefs, RE, Mundy, NI 2001The molecular basis of an avian plumage polymorphism in the wild: a melanocortin-1-receptor point mutation is perfectly associated with the melanic plumage morph of the bananaquit, Coereba flaveola Curr Biol11550557CrossRefPubMedGoogle Scholar
  46. Vage, DI, Lu, DS, Klungland, H, Lien, S, Adalsteinsson, S,  et al. 1997A non-epistatic interaction of agouti and extension in the fox, Vulpes vulpes Na Genet15311315CrossRefGoogle Scholar
  47. Vage, DI, Klungland, H, Lu, D, Cone, RD 1999Molecular and pharmacological characterization of dominant black coat color in sheepMamm Genome103943CrossRefPubMedGoogle Scholar
  48. Vila, C, Maldonado, JE, Wayne, RK 1999Phylogenetic relationships, evolution, and genetic diversity of the domestic dogJ Hered907177CrossRefPubMedGoogle Scholar
  49. Vrieling, H, Duhl, DM, Millar, SE, Miller, KA, Barsh, GS 1994Differences in dorsal and ventral pigmentation result from regional expression of the mouse agouti geneProc Natl Acad Sci USA9156675671PubMedGoogle Scholar
  50. Wayne, RK, Ostrander, EA 1999Origin, genetic diversity, and genome structure of the domestic dogBioessays21247257CrossRefPubMedGoogle Scholar
  51. Willard, DH, Bodnar, W, Harris, C, Kiefer, L, Nichols, JS,  et al. 1995Agouti structure and function: characterization of a potent alpha-melanocyte stimulating hormone receptor antagonistBiochemistry341234112346PubMedGoogle Scholar
  52. Willis, MB 1989Genetics of the dogHowellNew YorkGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Julie A. Kerns
    • 1
    • 5
  • J. Newton
    • 1
  • Tom G. Berryere
    • 2
  • Edward M. Rubin
    • 3
  • Jan-Fang Cheng
    • 3
  • Sheila M. Schmutz
    • 2
  • Gregory S. Barsh
    • 1
    • 4
    Email author
  1. 1.Departments of Genetics and PediatricsStanford UniversityStanfordUSA
  2. 2.Department of Animal and Poultry SciencesUniversity of SaskatchewanSaskatoonCanada
  3. 3.Genome Sciences DepartmentLawrence Berkeley National LaboratoryBerkeleyUSA
  4. 4.Beckman Center B271AStanford University School of MedicineStanfordUSA
  5. 5. Fred Hutchinson Cancer Research CenterSeattle

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