Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1996 Jul 1;15(13):3338–3350.

Characterization of a new melanocyte-specific gene (QNR-71) expressed in v-myc-transformed quail neuroretina.

N Turque 1, F Denhez 1, P Martin 1, N Planque 1, M Bailly 1, A Bègue 1, D Stéhelin 1, S Saule 1
PMCID: PMC451897  PMID: 8670835

Abstract

Quail neuroretina cells (QNR) infected with the v-myc-expressing retrovirus MC29 become pigmented after several passages in vitro. After differential screening of a cDNA library constructed from these cells, we have isolated a cDNA clone (QNR-71) which identifies an RNA expressed only in the pigmented layer of the retina and in the epidermis. This gene can also be induced in other cell types transformed by MC29, suggesting that QNR-71 may be regulated by the v-myc protein. Sequence analysis showed that the QNR-71 cDNA exhibits stretches of homologies with melanosomal proteins encoding genes. From bacterially expressed QNR-71 peptides we obtained rabbit antisera able to specifically recognize two proteins of 95 and 100 kDa in pigmented retinal cells, but not in the neuroretina. To study the regulation of QNR-71, we used promoter fragments linked to the CAT reporter gene, in transient co-expression assay. We observed an increase in CAT expression with a c-MYC and microphtalmia (mi) expression vectors. Both MYC and mi activate the QNR-71 promoter through direct binding to a CATGTG site present in the promoter fragment.

Full text

PDF
3338

Images in this article

3340Image
on p.3340
3341Image
on p.3341
3341Image
on p.3341
3342Image
on p.3342
3342Image
on p.3342
3343Image
on p.3343
3343Image
on p.3343
3344Image
on p.3344
3345Image
on p.3345
3347Image
on p.3347

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agata K., Kobayashi H., Itoh Y., Mochii M., Sawada K., Eguchi G. Genetic characterization of the multipotent dedifferentiated state of pigmented epithelial cells in vitro. Development. 1993 Aug;118(4):1025–1030. doi: 10.1242/dev.118.4.1025. [DOI] [PubMed] [Google Scholar]
  2. Alema S., Tato F., Boettiger D. myc and src oncogenes have complementary effects on cell proliferation and expression of specific extracellular matrix components in definitive chondroblasts. Mol Cell Biol. 1985 Mar;5(3):538–544. doi: 10.1128/mcb.5.3.538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Amati B., Brooks M. W., Levy N., Littlewood T. D., Evan G. I., Land H. Oncogenic activity of the c-Myc protein requires dimerization with Max. Cell. 1993 Jan 29;72(2):233–245. doi: 10.1016/0092-8674(93)90663-b. [DOI] [PubMed] [Google Scholar]
  4. Bello-Fernandez C., Packham G., Cleveland J. L. The ornithine decarboxylase gene is a transcriptional target of c-Myc. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7804–7808. doi: 10.1073/pnas.90.16.7804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bentley D. L., Groudine M. A block to elongation is largely responsible for decreased transcription of c-myc in differentiated HL60 cells. Nature. 1986 Jun 12;321(6071):702–706. doi: 10.1038/321702a0. [DOI] [PubMed] [Google Scholar]
  6. Bentley N. J., Eisen T., Goding C. R. Melanocyte-specific expression of the human tyrosinase promoter: activation by the microphthalmia gene product and role of the initiator. Mol Cell Biol. 1994 Dec;14(12):7996–8006. doi: 10.1128/mcb.14.12.7996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Benvenisty N., Leder A., Kuo A., Leder P. An embryonically expressed gene is a target for c-Myc regulation via the c-Myc-binding sequence. Genes Dev. 1992 Dec;6(12B):2513–2523. doi: 10.1101/gad.6.12b.2513. [DOI] [PubMed] [Google Scholar]
  8. Beug H., von Kirchbach A., Döderlein G., Conscience J. F., Graf T. Chicken hematopoietic cells transformed by seven strains of defective avian leukemia viruses display three distinct phenotypes of differentiation. Cell. 1979 Oct;18(2):375–390. doi: 10.1016/0092-8674(79)90057-6. [DOI] [PubMed] [Google Scholar]
  9. Bister K., Hayman M. J., Vogt P. K. Defectiveness of avian myelocytomatosis virus MC29: isolation of long-term nonproducer cultures and analysis of virus-specific polypeptide synthesis. Virology. 1977 Oct 15;82(2):431–448. doi: 10.1016/0042-6822(77)90017-4. [DOI] [PubMed] [Google Scholar]
  10. Blackwell T. K., Huang J., Ma A., Kretzner L., Alt F. W., Eisenman R. N., Weintraub H. Binding of myc proteins to canonical and noncanonical DNA sequences. Mol Cell Biol. 1993 Sep;13(9):5216–5224. doi: 10.1128/mcb.13.9.5216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Blackwell T. K., Kretzner L., Blackwood E. M., Eisenman R. N., Weintraub H. Sequence-specific DNA binding by the c-Myc protein. Science. 1990 Nov 23;250(4984):1149–1151. doi: 10.1126/science.2251503. [DOI] [PubMed] [Google Scholar]
  12. Blackwood E. M., Eisenman R. N. Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc. Science. 1991 Mar 8;251(4998):1211–1217. doi: 10.1126/science.2006410. [DOI] [PubMed] [Google Scholar]
  13. Carriere C., Plaza S., Martin P., Quatannens B., Bailly M., Stehelin D., Saule S. Characterization of quail Pax-6 (Pax-QNR) proteins expressed in the neuroretina. Mol Cell Biol. 1993 Dec;13(12):7257–7266. doi: 10.1128/mcb.13.12.7257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  15. Coll J., Saule S., Martin P., Raes M. B., Lagrou C., Graf T., Beug H., Simon I. E., Stehelin D. The cellular oncogenes c-myc, c-myb and c-erb are transcribed in defined types of avian hematopoietic cells. Exp Cell Res. 1983 Nov;149(1):151–162. doi: 10.1016/0014-4827(83)90388-9. [DOI] [PubMed] [Google Scholar]
  16. Dang C. V., Barrett J., Villa-Garcia M., Resar L. M., Kato G. J., Fearon E. R. Intracellular leucine zipper interactions suggest c-Myc hetero-oligomerization. Mol Cell Biol. 1991 Feb;11(2):954–962. doi: 10.1128/mcb.11.2.954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Desbiens X., Quéva C., Jaffredo T., Stéhelin D., Vandenbunder B. The relationship between cell proliferation and the transcription of the nuclear oncogenes c-myc, c-myb and c-ets-1 during feather morphogenesis in the chick embryo. Development. 1991 Mar;111(3):699–713. doi: 10.1242/dev.111.3.699. [DOI] [PubMed] [Google Scholar]
  18. Erba H. P., Gunning P., Kedes L. Nucleotide sequence of the human gamma cytoskeletal actin mRNA: anomalous evolution of vertebrate non-muscle actin genes. Nucleic Acids Res. 1986 Jul 11;14(13):5275–5294. doi: 10.1093/nar/14.13.5275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Falcone G., Tatò F., Alemà S. Distinctive effects of the viral oncogenes myc, erb, fps, and src on the differentiation program of quail myogenic cells. Proc Natl Acad Sci U S A. 1985 Jan;82(2):426–430. doi: 10.1073/pnas.82.2.426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Fauquet M., Stehelin D., Saule S. myc products induce the expression of catecholaminergic traits in quail neural crest-derived cells. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1546–1550. doi: 10.1073/pnas.87.4.1546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Fuerstenberg S., Beug H., Introna M., Khazaie K., Muñoz A., Ness S., Nordström K., Sap J., Stanley I., Zenke M. Ectopic expression of the erythrocyte band 3 anion exchange protein, using a new avian retrovirus vector. J Virol. 1990 Dec;64(12):5891–5902. doi: 10.1128/jvi.64.12.5891-5902.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hodgkinson C. A., Moore K. J., Nakayama A., Steingrímsson E., Copeland N. G., Jenkins N. A., Arnheiter H. Mutations at the mouse microphthalmia locus are associated with defects in a gene encoding a novel basic-helix-loop-helix-zipper protein. Cell. 1993 Jul 30;74(2):395–404. doi: 10.1016/0092-8674(93)90429-t. [DOI] [PubMed] [Google Scholar]
  24. Jaffredo T., Vandenbunder B., Dieterlen-Lièvre F. In situ study of c-myc protein expression during avian development. Development. 1989 Apr;105(4):679–695. doi: 10.1242/dev.105.4.679. [DOI] [PubMed] [Google Scholar]
  25. Kadesch T. Consequences of heteromeric interactions among helix-loop-helix proteins. Cell Growth Differ. 1993 Jan;4(1):49–55. [PubMed] [Google Scholar]
  26. Kato G. J., Barrett J., Villa-Garcia M., Dang C. V. An amino-terminal c-myc domain required for neoplastic transformation activates transcription. Mol Cell Biol. 1990 Nov;10(11):5914–5920. doi: 10.1128/mcb.10.11.5914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kawakami Y., Eliyahu S., Delgado C. H., Robbins P. F., Sakaguchi K., Appella E., Yannelli J. R., Adema G. J., Miki T., Rosenberg S. A. Identification of a human melanoma antigen recognized by tumor-infiltrating lymphocytes associated with in vivo tumor rejection. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6458–6462. doi: 10.1073/pnas.91.14.6458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kobayashi T., Urabe K., Orlow S. J., Higashi K., Imokawa G., Kwon B. S., Potterf B., Hearing V. J. The Pmel 17/silver locus protein. Characterization and investigation of its melanogenic function. J Biol Chem. 1994 Nov 18;269(46):29198–29205. [PubMed] [Google Scholar]
  29. Kozak M. Downstream secondary structure facilitates recognition of initiator codons by eukaryotic ribosomes. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8301–8305. doi: 10.1073/pnas.87.21.8301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kretzner L., Blackwood E. M., Eisenman R. N. Myc and Max proteins possess distinct transcriptional activities. Nature. 1992 Oct 1;359(6394):426–429. doi: 10.1038/359426a0. [DOI] [PubMed] [Google Scholar]
  31. Kwon B. S., Chintamaneni C., Kozak C. A., Copeland N. G., Gilbert D. J., Jenkins N., Barton D., Francke U., Kobayashi Y., Kim K. K. A melanocyte-specific gene, Pmel 17, maps near the silver coat color locus on mouse chromosome 10 and is in a syntenic region on human chromosome 12. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9228–9232. doi: 10.1073/pnas.88.20.9228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Landschulz W. H., Johnson P. F., McKnight S. L. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins. Science. 1988 Jun 24;240(4860):1759–1764. doi: 10.1126/science.3289117. [DOI] [PubMed] [Google Scholar]
  33. Langlois A. J., Ishizaki R., Beaudreau G. S., Kummer J. F., Beard J. W., Bolognesi D. P. Virus-infected avian cell lines established in vitro. Cancer Res. 1976 Nov;36(11 Pt 1):3894–3904. [PubMed] [Google Scholar]
  34. Lüscher B., Eisenman R. N. New light on Myc and Myb. Part I. Myc. Genes Dev. 1990 Dec;4(12A):2025–2035. doi: 10.1101/gad.4.12a.2025. [DOI] [PubMed] [Google Scholar]
  35. Martin P., Carriere C., Dozier C., Quatannens B., Mirabel M. A., Vandenbunder B., Stehelin D., Saule S. Characterization of a paired box- and homeobox-containing quail gene (Pax-QNR) expressed in the neuroretina. Oncogene. 1992 Sep;7(9):1721–1728. [PubMed] [Google Scholar]
  36. Martin P., Henry C., Ferre F., Duterque-Coquillaud M., Lagrou C., Ghysdael J., Debuire B., Stehelin D., Saule S. Transformation of quail embryo fibroblasts by a retrovirus carrying a normal human c-myc gene. EMBO J. 1986 Jul;5(7):1529–1533. doi: 10.1002/j.1460-2075.1986.tb04393.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Mochii M., Agata K., Eguchi G. Complete sequence and expression of a cDNA encoding a chicken 115-kDa melanosomal matrix protein. Pigment Cell Res. 1991 Feb;4(1):41–47. doi: 10.1111/j.1600-0749.1991.tb00312.x. [DOI] [PubMed] [Google Scholar]
  38. Murre C., McCaw P. S., Baltimore D. A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD, and myc proteins. Cell. 1989 Mar 10;56(5):777–783. doi: 10.1016/0092-8674(89)90682-x. [DOI] [PubMed] [Google Scholar]
  39. Park C. M., Hollenberg M. J. Basic fibroblast growth factor induces retinal regeneration in vivo. Dev Biol. 1989 Jul;134(1):201–205. doi: 10.1016/0012-1606(89)90089-4. [DOI] [PubMed] [Google Scholar]
  40. Pittack C., Jones M., Reh T. A. Basic fibroblast growth factor induces retinal pigment epithelium to generate neural retina in vitro. Development. 1991 Oct;113(2):577–588. doi: 10.1242/dev.113.2.577. [DOI] [PubMed] [Google Scholar]
  41. Plaza S., Dozier C., Saule S. Quail Pax-6 (Pax-QNR) encodes a transcription factor able to bind and trans-activate its own promoter. Cell Growth Differ. 1993 Dec;4(12):1041–1050. [PubMed] [Google Scholar]
  42. Plaza S., Dozier C., Turque N., Saule S. Quail Pax-6 (Pax-QNR) mRNAs are expressed from two promoters used differentially during retina development and neuronal differentiation. Mol Cell Biol. 1995 Jun;15(6):3344–3353. doi: 10.1128/mcb.15.6.3344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Prendergast G. C., Ziff E. B. DNA-binding motif. Nature. 1989 Oct 5;341(6241):392–392. doi: 10.1038/341392a0. [DOI] [PubMed] [Google Scholar]
  44. Reisman D., Elkind N. B., Roy B., Beamon J., Rotter V. c-Myc trans-activates the p53 promoter through a required downstream CACGTG motif. Cell Growth Differ. 1993 Feb;4(2):57–65. [PubMed] [Google Scholar]
  45. Robbins L. S., Nadeau J. H., Johnson K. R., Kelly M. A., Roselli-Rehfuss L., Baack E., Mountjoy K. G., Cone R. D. Pigmentation phenotypes of variant extension locus alleles result from point mutations that alter MSH receptor function. Cell. 1993 Mar 26;72(6):827–834. doi: 10.1016/0092-8674(93)90572-8. [DOI] [PubMed] [Google Scholar]
  46. Steel K. P., Davidson D. R., Jackson I. J. TRP-2/DT, a new early melanoblast marker, shows that steel growth factor (c-kit ligand) is a survival factor. Development. 1992 Aug;115(4):1111–1119. doi: 10.1242/dev.115.4.1111. [DOI] [PubMed] [Google Scholar]
  47. Turque N., Plaza S., Radvanyi F., Carriere C., Saule S. Pax-QNR/Pax-6, a paired box- and homeobox-containing gene expressed in neurons, is also expressed in pancreatic endocrine cells. Mol Endocrinol. 1994 Jul;8(7):929–938. doi: 10.1210/mend.8.7.7984154. [DOI] [PubMed] [Google Scholar]
  48. Weterman M. A., Ajubi N., van Dinter I. M., Degen W. G., van Muijen G. N., Ruitter D. J., Bloemers H. P. nmb, a novel gene, is expressed in low-metastatic human melanoma cell lines and xenografts. Int J Cancer. 1995 Jan 3;60(1):73–81. doi: 10.1002/ijc.2910600111. [DOI] [PubMed] [Google Scholar]
  49. Yasumoto K., Yokoyama K., Shibata K., Tomita Y., Shibahara S. Microphthalmia-associated transcription factor as a regulator for melanocyte-specific transcription of the human tyrosinase gene. Mol Cell Biol. 1994 Dec;14(12):8058–8070. doi: 10.1128/mcb.14.12.8058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Yavuzer U., Keenan E., Lowings P., Vachtenheim J., Currie G., Goding C. R. The Microphthalmia gene product interacts with the retinoblastoma protein in vitro and is a target for deregulation of melanocyte-specific transcription. Oncogene. 1995 Jan 5;10(1):123–134. [PubMed] [Google Scholar]
  51. Zhou B. K., Kobayashi T., Donatien P. D., Bennett D. C., Hearing V. J., Orlow S. J. Identification of a melanosomal matrix protein encoded by the murine si (silver) locus using "organelle scanning". Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7076–7080. doi: 10.1073/pnas.91.15.7076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. doi: 10.1111/j.1432-1033.1983.tb07424.x. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES