, Volume 108, Issue 3, pp 239–252 | Cite as

Sub-cellular Localisation of the White/Scarlet ABC Transporter to Pigment Granule Membranes Within the Compound Eye of Drosophila Melanogaster

  • Susan M. Mackenzie
  • Antony J. Howells
  • Graeme B. Cox
  • Gary D. Ewart


The white, scarlet, and browngenes of Drosophila melanogasterencode ABC transporters involved with the uptake and storage of metabolic precursors to the red and brown eye colour pigments. It has generally been assumed that these proteins are localised in the plasma membrane and transport precursor molecules from the heamolymph into the eye pigment cells. However, the immuno-electron microscopy experiments in this study reveal that the White and Scarlet proteins are located in the membranes of pigment granules within pigment cells and retinula cells of the compound eye. No evidence of their presence in the plasma membrane was observed. This result suggests that, rather than tranporting tryptophan into the cell across the plasma membrane, the White/Scarlet complex transports a metabolic intermediate (such as 3-hydroxy kynurenine) from the cytoplasm into the pigment granules. Other functional implications of this new finding are discussed.

ABC transporter transport ATPase whitegene scarletgene browngene pigment precursor transport 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bingham, P.M., R. Levis & G.M. Rubin, 1981. Cloning of DNA sequences from the whitelocus of Drosophila melanogasterby a novel and general method. Cell 25: 693–704.PubMedCrossRefGoogle Scholar
  2. Cagan, R.L. & D.F. Ready, 1989. The emergence of order in the Drosophilapupal retina. Develop. Biol. 136: 346–362.PubMedCrossRefGoogle Scholar
  3. Castagna, M., C. Shayakul, D. Trotti, V.F. Sacchi, W.R. Harvey & M.A. Hediger, 1997. Molecular characteristics of mammalian and insect amino acid transporters: implications for amino acid homeostasis. J. Exp. Biol. 200: 269–286.PubMedGoogle Scholar
  4. Chen, H., C. Rossier, M.D. Lalioti, A. Lynn, A. Chakravarti, G. Perrin & S.E. Antonarakis, 1996. Cloning of the cDNA for a human homologue of the DrosophilaWhite gene and mapping to chromosome 21q22.3. Amer. J. Hum. Genet. 59: 66–75.PubMedGoogle Scholar
  5. Croop, J.M., 1998. Evolutionary relationships among ABC transporters. Meth. Enzymol. 292: 101–116.PubMedGoogle Scholar
  6. Dreeson, T.D., D.H. Johnson & S. Henikoff, 1988. The Brown protein of Drosophila melanogasteris similar to the White protein and to components of active transport complexes. Mol. Cell. Biol. 8: 5206–5215.Google Scholar
  7. Ewart, G.D., D. Cannell, G.B. Cox & A.J. Howells, 1994. Mutational analysis of the traffic ATPase (ABC) transporters involved in uptake of eye pigment precursors in Drosophila melanogaster. J. Biol. Chem. 269: 10370–10377.PubMedGoogle Scholar
  8. Ewart, G.D. & A.J. Howells, 1998. ABC transporters involved in transport of eye pigment precursors in Drosophila melanogaster. Meth. Enzymol. 292: 213–224.PubMedGoogle Scholar
  9. Fjose, A., L.C. Polito, U. Weber & W.J. Gehring 1984. Developmental expression of the whitelocus of Drosophila melanogaster. EMBO J. 3: 2087–2094.PubMedGoogle Scholar
  10. Forrest, H.S., 1957. Pigment Cell Biology. Academic Press, New York.Google Scholar
  11. Harlow, E. & D. Lane, 1988. Antibodies, A Laboratory Manual. Cold Spring Harbour Laboratory, New York.Google Scholar
  12. Higgins, C.F., 1992. ABC transporters: frommicrooganisms toman. Ann. Rev. Cell Biol. 8: 67–113.PubMedGoogle Scholar
  13. Höning, S., I.V. Sandoval & Figura, K.V., 1998. A di-leucine-based motif in the cytoplasmic tail of LIMP-II and tyrosinase mediates selective binding of AP-3. EMBO J. 17: 1304–1314.PubMedCrossRefGoogle Scholar
  14. Howells, A.J. & R.L. Ryall, 1975. A biochemical study of the Scarleteye-color mutant of Drosophila melanogaster. Bioch Genet. 13: 273–282.CrossRefGoogle Scholar
  15. Howells, A.J., K.M. Summers & R.L. Ryall, 1977. Developmental patterns of 3-hydroxykynurenine accumulation in whiteand various other eye color mutants of Drosophila melanogaster. Biochem. Genetics 15: 1049–1059.CrossRefGoogle Scholar
  16. King, L.A. & R.D. Possee, 1992. The Baculovirus Expresssion System, A laboratory Guide. Chapman & Hall, London.Google Scholar
  17. Kurkulos, M., J.M. Weinberg, M.E. Pepling & S.M. Mount, 1991. Polyadenylation in copiarequires unusually distant upstream sequences. Proc. Nat. Acad. Sci. USA 88: 3038–3042.PubMedCrossRefGoogle Scholar
  18. Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.PubMedCrossRefGoogle Scholar
  19. Levis, R., T. Hazelrigg & G.M. Rubin, 1985a. Effects of genomic position on the expression of transduced copies of the whitegene of Drosophila. Science 229: 558–561.PubMedGoogle Scholar
  20. Levis, R., T. Hazelrigg & G.M. Rubin, 1985b. Separable cis-acting control elements for expression of theWhite gene of Drosophila. EMBO J. 4: 3489–3499.PubMedGoogle Scholar
  21. Linzen, B., 1974. The tryptophan to ommochrome pathway in insects. Adv. Insect Physiol. 10: 117–245.Google Scholar
  22. Lloyd, V., M. Ramaswami & Krämer, H. 1998. Not just pretty eyes: Drosophilaeye-colour mutations and lysosomal delivery. Trends Cell Biol. 8: 257–259.PubMedCrossRefGoogle Scholar
  23. Lu, Y.-A., P. Clavijo, M. Galantino, Z.-Y. Shen, W. Liu & J.P. Tam, 1991. Chemically unambiguous peptide immunogen: preparation, orientation and antigenicity of purified peptide conjugated to the multiple antigen peptide system. Mol. Immunol. 28: 623–630.PubMedCrossRefGoogle Scholar
  24. Mackenzie, S., M.R. Brooker, T.R. Gill, G.B. Cox, A.J. Howells & G.D. Ewart, 1999. Mutations in the whitegene of Drosophila melanogasteraffecting ABC transporters that determine eye colouration. Biochim. et Biophys. Acta 1419: 173–185.CrossRefGoogle Scholar
  25. McGivan, J.D. & M. Pastor-Anglada, 1994. Regulatory and molecular aspects of mammalian amino acid transport. Biochem. J. 299: 321–334.PubMedGoogle Scholar
  26. Odorizzi, G., C.R. Cowles & S.D. Emr, 1998. The AP-3 complex: a coat of many colours. Trends Cell Biol. 8: 282–288.PubMedCrossRefGoogle Scholar
  27. O'Hare, K., C. Murphy, R. Levis & G.M. Rubin, 1984. DNA Sequence of the whitelocus of Drosophila melanogaster. J. Mol. Biol. 180: 437–455.PubMedCrossRefGoogle Scholar
  28. Ooi, C.E., J.E. Moreira, E.C. Dell'Angelica, G. Poy, D.A. Wassarman & J.S. Bonifacino, 1997. Altered expression of a novel adaptin leads to defective pigment granule biogenesis in the Drosophilaeye colour mutant garnet. EMBO J. 16: 4508–4518.PubMedCrossRefGoogle Scholar
  29. Pepling, M. & S.M. Mount, 1990. Sequence of a cDNA from the Drosophila melanogaster whitegene. Nucleic Acids Res. 18: 1633.PubMedGoogle Scholar
  30. Phillips, J.P. & H.S. Forrest, 1980. Ommochromes and pteridines pp. 542–611, in The Genetics and Biology of Drosophila, edited by M. Ashburner & T.R.F. Wright. Vol. 2d, Academic Press, New York and London.Google Scholar
  31. Potterf, S.B., J. Muler, I. Bernardini, F. Tietze, T. Kobayashi, V.J. Hearing & W.A. Gahl, 1996. Characterization of a melanosomal transport system in murine melanocytes mediating entry of the melanogenic substrate tyrosine. J. Biol. Chem. 271: 4002–4008.PubMedCrossRefGoogle Scholar
  32. Refranz, P. & S. Benzer, 1989. Monoclonal antibody probes discriminate early and late mutant defects in development of the Drosophilaretina. Develop. Biol. 136: 411–429.CrossRefGoogle Scholar
  33. Roberts, D.B., 1986. Drosophila: A Practical Approach. IRL Press Limited, Oxford.Google Scholar
  34. Ryall, R.L. & A.J. Howells, 1974. Ommochrome biosynthetic pathway of Drosophila melanogaster: Variations in the levels of enzyme activities and intermediates during adult development. Insect Biochem. 6: 135–142.CrossRefGoogle Scholar
  35. Sambrook, J., E.F. Fritsch & T. Maniatis, 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
  36. Savary, S., F. Denizot, M.-F. Luciani, M.-G. Mattei & G. Chimini, 1996. Molecular cloning of a mammalian ABC transporter homologous to Drosophila whitegene. Mammal. Genome 7: 673–676.Google Scholar
  37. Schraermeyer, U. & M. Dohms, 1993. Atypical granules in the eyes of whitemutant Drosophila melanogasterare lysosome-related organelles. Pigment Cell Res. 6: 73–84.PubMedGoogle Scholar
  38. Shotwell, M.A. & D.L. Oxender, 1983. The regulation of neutral amino acid transport by amino acid availability in animal cells. Trends Biochem. Sci. 8: 314–316.CrossRefGoogle Scholar
  39. Shoup, J.R., 1966. The development of pigment granules in the eyes of wild type and mutant Drosophila melanogaster. J. Cell Biol. 29: 223–249.PubMedCrossRefGoogle Scholar
  40. Simpson, F., A.A. Peden, L. Christopoulou & M.S. Robinson, 1997. Characterisation of the Adaptor-related protein complex, AP-3. J. Cell Biol. 137: 835–845.PubMedCrossRefGoogle Scholar
  41. Stark, W.S. & R. Sapp, 1988. Eye color pigment granules in wildtype and mutant Drosophila melanogaster. Canadian J. Zool. 66: 1301–1308.CrossRefGoogle Scholar
  42. Sullivan, D.T., L.A. Bell, D.R. Paton & M.C. Sullivan, 1979. Purine transport by malpighian tubules of pteridine-deficient eye color mutants of Drosophila melanogaster. Biochem. Genet. 17: 565–573.PubMedCrossRefGoogle Scholar
  43. Sullivan, D.T., L.A. Bell, D.R. Paton & M.C. Sullivan, 1980. Genetic and functional analysis of tryptophan transport in malpighian tubules of Drosophila. Biochem. Genet. 18: 1109–1131.PubMedCrossRefGoogle Scholar
  44. Sullivan, D.T. & M.C. Sullivan, 1975. Transport defects as the physiological basis for eye color mutants of Drosophila melanogaster. Biochem. Genet. 13: 603–613.PubMedCrossRefGoogle Scholar
  45. Summers, K.M., A.J. Howells & N.A. Pyliotis, 1982. Biology of eye pigmentation in insects. Adv. Insect Physiol. 16: 119–166.CrossRefGoogle Scholar
  46. Tearle, R.G., J.M. Belote, M. McKeown, B.S. Baker & A.J. Howells, 1989. Cloning and characterisation of the Scarletgene of Drosophila melanogaster. Genet. 122: 595–606.Google Scholar
  47. Towbin, H., T. Staehelin & J. Gordon, 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Nat. Acad. Sci. USA 76: 4350–4354.PubMedCrossRefGoogle Scholar
  48. Venkateswaran, A., J.J. Repa, J.A. Lobaccaro, A. Bronson, D.J. Mangelsdorf & P.A. Edwards, 2000. Human White/murine ABC8 mRNA levels are highly induced in lipid-loaded macrophages. J. Biol. Chem. 275: 14700–14707.PubMedCrossRefGoogle Scholar
  49. Wiley, K. & H.S. Forrest, 1981. Terminal synthesis of xanthommatin in Drosophila melanogaster. IV. enzymatic and nonenzymatic catalysis. Biochem. Genet. 19: 1211–1221.PubMedCrossRefGoogle Scholar
  50. Yagi, S. & H. Ogawa, 1996. Effect of tryptophan metabolites on fluorescent granules in the malpighian tubules of eye color mutants of Drosophila melanogaster. Zool. Sci. 13: 97–104.PubMedCrossRefGoogle Scholar
  51. Zwiebel, L.J., G. Saccone, A. Zacharopoulou, N.J. Besansky, G. Favia, F.H. Collins, C. Louis & F.C. Kafatos, 1995. The whitegene of Ceratitis capitata: a phenotypic marker for germline transformation. Science 270: 2005–2007.PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Susan M. Mackenzie
    • 1
  • Antony J. Howells
    • 2
  • Graeme B. Cox
    • 1
  • Gary D. Ewart
    • 3
  1. 1.Division of Biochemistry and Molecular Biology, John Curtin School of Medical ResearchThe Australian National UniversityCanberra CityAustralia
  2. 2.Division of Biochemistry and Molecular Biology, Faculty of ScienceThe Australian National UniversityCanberra CityAustralia
  3. 3.Division of Biochemistry and Molecular Biology, John Curtin School of Medical ResearchThe Australian National UniversityCanberra CityAustralia

Personalised recommendations