Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1992 Feb 1;116(3):683–693. doi: 10.1083/jcb.116.3.683

Differential localizations of and requirements for the two Drosophila ninaC kinase/myosins in photoreceptor cells

PMCID: PMC2289314  PMID: 1730774

Abstract

The ninaC gene encodes two retinal specific proteins (p132 and p174) consisting of a protein kinase domain joined to a domain homologous to the head region of the myosin heavy chain. The putative myosin domain of p174 is linked at the COOH-terminus to a tail which has some similarities to myosin-I tails. In the current report, we demonstrate that the ninaC mutation results in light- and age-dependent retinal degeneration. We also show that ninaC flies display an electrophysiological phenotype before any discernible retinal degeneration indicating that the electrophysiological defect is the primary effect of the mutation. This suggests that ninaC has a role in phototransduction and that the retinal degeneration is a secondary effect resulting from the defect in phototransduction. To examine the requirements for the individual ninaC isoforms, mutant alleles were generated which express only p132 or p174. Elimination of p174 resulted in a ninaC phenotype as strong as the null allele; however, elimination of p132 had little if any effect. As a first step in investigating the basis for the difference in requirements for p174 and p132 we performed immuno-localization at the electron microscopic level and found that the two isoforms display different subcellular distributions in the photoreceptor cells. The p132 protein is restricted primarily to the cytoplasm and p174 to the rhabdomeres, the microvillar structure which is the site of action of many of the steps in phototransduction. This suggests that the p174 myosin-I type tail is the domain responsible for association with the rhabdomeres and that the substrate for the p174 putative kinase may be a rhabdomeric protein important in photo- transduction.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Adams R. J., Pollard T. D. Propulsion of organelles isolated from Acanthamoeba along actin filaments by myosin-I. Nature. 1986 Aug 21;322(6081):754–756. doi: 10.1038/322754a0. [DOI] [PubMed] [Google Scholar]
  2. Aho A. C., Donner K., Hydén C., Larsen L. O., Reuter T. Low retinal noise in animals with low body temperature allows high visual sensitivity. Nature. 1988 Jul 28;334(6180):348–350. doi: 10.1038/334348a0. [DOI] [PubMed] [Google Scholar]
  3. Arikawa K., Hicks J. L., Williams D. S. Identification of actin filaments in the rhabdomeral microvilli of Drosophila photoreceptors. J Cell Biol. 1990 Jun;110(6):1993–1998. doi: 10.1083/jcb.110.6.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blackshear P. J., Nairn A. C., Kuo J. F. Protein kinases 1988: a current perspective. FASEB J. 1988 Nov;2(14):2957–2969. doi: 10.1096/fasebj.2.14.2972578. [DOI] [PubMed] [Google Scholar]
  5. Bloomquist B. T., Shortridge R. D., Schneuwly S., Perdew M., Montell C., Steller H., Rubin G., Pak W. L. Isolation of a putative phospholipase C gene of Drosophila, norpA, and its role in phototransduction. Cell. 1988 Aug 26;54(5):723–733. doi: 10.1016/s0092-8674(88)80017-5. [DOI] [PubMed] [Google Scholar]
  6. Bowes C., Li T., Danciger M., Baxter L. C., Applebury M. L., Farber D. B. Retinal degeneration in the rd mouse is caused by a defect in the beta subunit of rod cGMP-phosphodiesterase. Nature. 1990 Oct 18;347(6294):677–680. doi: 10.1038/347677a0. [DOI] [PubMed] [Google Scholar]
  7. Dryja T. P., McGee T. L., Reichel E., Hahn L. B., Cowley G. S., Yandell D. W., Sandberg M. A., Berson E. L. A point mutation of the rhodopsin gene in one form of retinitis pigmentosa. Nature. 1990 Jan 25;343(6256):364–366. doi: 10.1038/343364a0. [DOI] [PubMed] [Google Scholar]
  8. Farber J. L. The role of calcium in cell death. Life Sci. 1981 Sep 28;29(13):1289–1295. doi: 10.1016/0024-3205(81)90670-6. [DOI] [PubMed] [Google Scholar]
  9. Hanks S. K., Quinn A. M., Hunter T. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science. 1988 Jul 1;241(4861):42–52. doi: 10.1126/science.3291115. [DOI] [PubMed] [Google Scholar]
  10. Harris W. A., Stark W. S. Hereditary retinal degeneration in Drosophila melanogaster. A mutant defect associated with the phototransduction process. J Gen Physiol. 1977 Mar;69(3):261–291. doi: 10.1085/jgp.69.3.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hayden S. M., Wolenski J. S., Mooseker M. S. Binding of brush border myosin I to phospholipid vesicles. J Cell Biol. 1990 Aug;111(2):443–451. doi: 10.1083/jcb.111.2.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hillman P., Hochstein S., Minke B. A visual pigment with two physiologically active stable states. Science. 1972 Mar 31;175(4029):1486–1488. doi: 10.1126/science.175.4029.1486. [DOI] [PubMed] [Google Scholar]
  13. Hotta Y., Benzer S. Genetic dissection of the Drosophila nervous system by means of mosaics. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1156–1163. doi: 10.1073/pnas.67.3.1156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Inoue H., Yoshioka T., Hotta Y. Diacylglycerol kinase defect in a Drosophila retinal degeneration mutant rdgA. J Biol Chem. 1989 Apr 5;264(10):5996–6000. [PubMed] [Google Scholar]
  15. Korn E. D., Hammer J. A., 3rd Myosins of nonmuscle cells. Annu Rev Biophys Biophys Chem. 1988;17:23–45. doi: 10.1146/annurev.bb.17.060188.000323. [DOI] [PubMed] [Google Scholar]
  16. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  17. Matsumoto H., Isono K., Pye Q., Pak W. L. Gene encoding cytoskeletal proteins in Drosophila rhabdomeres. Proc Natl Acad Sci U S A. 1987 Feb;84(4):985–989. doi: 10.1073/pnas.84.4.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McClary J. A., Witney F., Geisselsoder J. Efficient site-directed in vitro mutagenesis using phagemid vectors. Biotechniques. 1989 Mar;7(3):282–289. [PubMed] [Google Scholar]
  19. Meyertholen E. P., Stein P. J., Williams M. A., Ostroy S. E. Studies of the Drosophila norpA phototransduction mutant. II. Photoreceptor degeneration and rhodopsin maintenance. J Comp Physiol A. 1987 Nov;161(6):793–798. doi: 10.1007/BF00610221. [DOI] [PubMed] [Google Scholar]
  20. Mismer D., Michael W. M., Laverty T. R., Rubin G. M. Analysis of the promoter of the Rh2 opsin gene in Drosophila melanogaster. Genetics. 1988 Sep;120(1):173–180. doi: 10.1093/genetics/120.1.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Montell C., Rubin G. M. The Drosophila ninaC locus encodes two photoreceptor cell specific proteins with domains homologous to protein kinases and the myosin heavy chain head. Cell. 1988 Mar 11;52(5):757–772. doi: 10.1016/0092-8674(88)90413-8. [DOI] [PubMed] [Google Scholar]
  22. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pollard T. D., Doberstein S. K., Zot H. G. Myosin-I. Annu Rev Physiol. 1991;53:653–681. doi: 10.1146/annurev.ph.53.030191.003253. [DOI] [PubMed] [Google Scholar]
  24. Rubin G. M., Spradling A. C. Genetic transformation of Drosophila with transposable element vectors. Science. 1982 Oct 22;218(4570):348–353. doi: 10.1126/science.6289436. [DOI] [PubMed] [Google Scholar]
  25. Rüther U., Müller-Hill B. Easy identification of cDNA clones. EMBO J. 1983;2(10):1791–1794. doi: 10.1002/j.1460-2075.1983.tb01659.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Smith D. P., Stamnes M. A., Zuker C. S. Signal transduction in the visual system of Drosophila. Annu Rev Cell Biol. 1991;7:161–190. doi: 10.1146/annurev.cb.07.110191.001113. [DOI] [PubMed] [Google Scholar]
  27. Spradling A. C., Rubin G. M. Transposition of cloned P elements into Drosophila germ line chromosomes. Science. 1982 Oct 22;218(4570):341–347. doi: 10.1126/science.6289435. [DOI] [PubMed] [Google Scholar]
  28. Srebro R., Behbehani M. The thermal origin of spontaneous activity in the Limulus photoreceptor. J Physiol. 1972 Jul;224(2):349–361. doi: 10.1113/jphysiol.1972.sp009899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Steele F., O'Tousa J. E. Rhodopsin activation causes retinal degeneration in Drosophila rdgC mutant. Neuron. 1990 Jun;4(6):883–890. doi: 10.1016/0896-6273(90)90141-2. [DOI] [PubMed] [Google Scholar]
  30. Thompson P., Findlay J. B. Phosphorylation of ovine rhodopsin. Identification of the phosphorylated sites. Biochem J. 1984 Jun 15;220(3):773–780. doi: 10.1042/bj2200773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Travis G. H., Sutcliffe J. G., Bok D. The retinal degeneration slow (rds) gene product is a photoreceptor disc membrane-associated glycoprotein. Neuron. 1991 Jan;6(1):61–70. doi: 10.1016/0896-6273(91)90122-g. [DOI] [PubMed] [Google Scholar]
  32. Vihtelic T. S., Hyde D. R., O'Tousa J. E. Isolation and characterization of the Drosophila retinal degeneration B (rdgB) gene. Genetics. 1991 Apr;127(4):761–768. doi: 10.1093/genetics/127.4.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Warrick H. M., Spudich J. A. Myosin structure and function in cell motility. Annu Rev Cell Biol. 1987;3:379–421. doi: 10.1146/annurev.cb.03.110187.002115. [DOI] [PubMed] [Google Scholar]
  34. Wilden U., Hall S. W., Kühn H. Phosphodiesterase activation by photoexcited rhodopsin is quenched when rhodopsin is phosphorylated and binds the intrinsic 48-kDa protein of rod outer segments. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1174–1178. doi: 10.1073/pnas.83.5.1174. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES