The RGS gene loco is essential for male reproductive system differentiation in Drosophila melanogaster (original) (raw)

Abstract

Background The loco gene encodes several different isoforms of a regulator of G-protein signalling. These different isoforms of LOCO are part of a pathway enabling cells to respond to external signals. LOCO is known to be required at various developmental stages including neuroblast division, glial cell formation and oogenesis. Less is known about LOCO and its involvement in male development therefore to gain further insight into the role of LOCO in development we carried out a genetic screen and analysed males with reduced fertility. Results We identified a number of lethal loco mutants and four semi-lethal lines, which generate males with reduced fertility. We have identified a fifth loco transcript and show that it is differentially expressed in developing pupae. We have characterised the expression pattern of all loco transcripts during pupal development in the adult testes, both in wild type and loco mutant strains. In addition we also show that there are various G-protein α su...

Loading...

Loading Preview

Sorry, preview is currently unavailable. You can download the paper by clicking the button above.

References (37)

  1. De Vries L, Mousli M, Wurmser A, Farquhar MG: GAIP, a protein that specifically interacts with the trimeric G protein G alpha i3, is a member of a protein family with a highly con- served core domain. Proc Natl Acad Sci U S A 1995, 92(25):11916-11920.
  2. Neubig RR, Siderovski DP: Regulators of G-protein signalling as new central nervous system drug targets. Nat Rev Drug Discov 2002, 1(3):187-197.
  3. Berman DM, Wilkie TM, Gilman AG: GAIP and RGS4 are GTPase-activating proteins for the Gi subfamily of G protein alpha subunits. Cell 1996, 86(3):445-452.
  4. Hunt TW, Fields TA, Casey PJ, Peralta EG: RGS10 is a selective activator of G alpha i GTPase activity. Nature 1996, 383(6596):175-177.
  5. Granderath S, Klambt C: Identification and functional analysis of the Drosophila gene loco. Methods Enzymol 2004, 389:350-363.
  6. Willert K, Logan CY, Arora A, Fish M, Nusse R: A Drosophila Axin homolog, Daxin, inhibits Wnt signaling. Development 1999, 126(18):4165-4173.
  7. Liu C, Li Y, Semenov M, Han C, Baeg GH, Tan Y, Zhang Z, Lin X, He X: Control of beta-catenin phosphorylation/degradation by a dual-kinase mechanism. Cell 2002, 108(6):837-847.
  8. Hart MJ, de los Santos R, Albert IN, Rubinfeld B, Polakis P: Down- regulation of beta-catenin by human Axin and its association with the APC tumor suppressor, beta-catenin and GSK3 beta. Curr Biol 1998, 8(10):573-581.
  9. Logan CY, Nusse R: The Wnt signaling pathway in develop- ment and disease. Annu Rev Cell Dev Biol 2004, 20:781-810.
  10. Castellone MD, Teramoto H, Williams BO, Druey KM, Gutkind JS: Prostaglandin E2 promotes colon cancer cell growth through a Gs-axin-beta-catenin signaling axis. Science 2005, 310(5753):1504-1510.
  11. Lannutti BJ, Schneider LE: Gprk2 controls cAMP levels in Dro- sophila development. Dev Biol 2001, 233(1):174-185.
  12. Schneider LE, Spradling AC: The Drosophila G-protein-coupled receptor kinase homologue Gprk2 is required for egg mor- phogenesis. Development 1997, 124(13):2591-2602.
  13. Yu F, Wang H, Qian H, Kaushik R, Bownes M, Yang X, Chia W: Loco- motion defects, together with Pins, regulates heterotrimeric G-protein signaling during Drosophila neuroblast asymmet- ric divisions. Genes Dev 2005, 19(11):1341-1353.
  14. Granderath S, Stollewerk A, Greig S, Goodman CS, O'Kane CJ, Klambt C: loco encodes an RGS protein required for Dro- sophila glial differentiation. Development 1999, 126(8):1781-1791.
  15. Pathirana S, Zhao D, Bownes M: The Drosophila RGS protein Loco is required for dorsal/ventral axis formation of the egg and embryo, and nurse cell dumping. Mech Dev 2001, 109(2):137-150.
  16. Wolfgang WJ, Quan F, Thambi N, Forte M: Restricted spatial and temporal expression of G-protein alpha subunits during Dro- sophila embryogenesis. Development 1991, 113(2):527-538.
  17. Provost NM, Somers DE, Hurley JB: A Drosophila melanogaster G protein alpha subunit gene is expressed primarily in embryos and pupae. J Biol Chem 1988, 263(24):12070-12076.
  18. Fremion F, Astier M, Zaffran S, Guillen A, Homburger V, Semeriva M: The heterotrimeric protein Go is required for the formation of heart epithelium in Drosophila. J Cell Biol 1999, 145(5):1063-1076.
  19. Thambi NC, Quan F, Wolfgang WJ, Spiegel A, Forte M: Immunolog- ical and molecular characterization of Go alpha-like proteins in the Drosophila central nervous system. J Biol Chem 1989, 264(31):18552-18560.
  20. Wolfgang WJ, Quan F, Goldsmith P, Unson C, Spiegel A, Forte M: Immunolocalization of G protein alpha-subunits in the Dro- sophila CNS. J Neurosci 1990, 10(3):1014-1024.
  21. Katanaev VL, Tomlinson A: Dual roles for the trimeric G protein Go in asymmetric cell division in Drosophila. Proc Natl Acad Sci U S A 2006, 103(17):6524-6529.
  22. de Sousa SM, Hoveland LL, Yarfitz S, Hurley JB: The Drosophila Go alpha-like G protein gene produces multiple transcripts and is expressed in the nervous system and in ovaries. J Biol Chem 1989, 264(31):18544-18551.
  23. Quan F, Wolfgang WJ, Forte M: A Drosophila G-protein alpha subunit, Gf alpha, expressed in a spatially and temporally restricted pattern during Drosophila development. Proc Natl Acad Sci U S A 1993, 90(9):4236-4240.
  24. Lee YJ, Dobbs MB, Verardi ML, Hyde DR: dgq: a drosophila gene encoding a visual system-specific G alpha molecule. Neuron 1990, 5(6):889-898.
  25. Strathmann M, Simon MI: G protein diversity: a distinct class of alpha subunits is present in vertebrates and invertebrates. Proc Natl Acad Sci U S A 1990, 87(23):9113-9117.
  26. Alvarez CE, Robison K, Gilbert W: Novel Gq alpha isoform is a candidate transducer of rhodopsin signaling in a Drosophila testes-autonomous pacemaker. Proc Natl Acad Sci U S A 1996, 93(22):12278-12282.
  27. Koelle MR, Horvitz HR: EGL-10 regulates G protein signaling in the C. elegans nervous system and shares a conserved domain with many mammalian proteins. Cell 1996, 84(1):115-125.
  28. Snow BE, Antonio L, Suggs S, Gutstein HB, Siderovski DP: Molecular cloning and expression analysis of rat Rgs12 and Rgs14. Bio- chem Biophys Res Commun 1997, 233(3):770-777.
  29. Snow BE, Hall RA, Krumins AM, Brothers GM, Bouchard D, Brothers CA, Chung S, Mangion J, Gilman AG, Lefkowitz RJ, Siderovski DP: GTPase activating specificity of RGS12 and binding specifi- city of an alternatively spliced PDZ (PSD-95/Dlg/ZO-1) domain. J Biol Chem 1998, 273(28):17749-17755.
  30. Snow BE, Krumins AM, Brothers GM, Lee SF, Wall MA, Chung S, Mangion J, Arya S, Gilman AG, Siderovski DP: A G protein gamma subunit-like domain shared between RGS11 and other RGS proteins specifies binding to Gbeta5 subunits. Proc Natl Acad Sci U S A 1998, 95(22):13307-13312.
  31. Thomas EA, Danielson PE, Sutcliffe JG: RGS9: a regulator of G- protein signalling with specific expression in rat and mouse striatum. J Neurosci Res 1998, 52(1):118-124.
  32. Wolfgang WJ, Forte M: Posterior localization of the Drosophila Gi alpha protein during early embryogenesis requires a sub- set of the posterior group genes. Int J Dev Biol 1995, 39(4):581-586.
  33. Deng W, Leaper K, Bownes M: A targeted gene silencing tech- nique shows that Drosophila myosin VI is required for egg chamber and imaginal disc morphogenesis. J Cell Sci 1999, 112 ( Pt 21):3677-3690.
  34. Schultz J, Milpetz F, Bork P, Ponting CP: SMART, a simple modu- lar architecture research tool: identification of signaling domains. Proc Natl Acad Sci U S A 1998, 95(11):5857-5864.
  35. Marchler-Bauer A, Anderson JB, Cherukuri PF, DeWeese-Scott C, Geer LY, Gwadz M, He S, Hurwitz DI, Jackson JD, Ke Z, Lanczycki CJ, Liebert CA, Liu C, Lu F, Marchler GH, Mullokandov M, Shoemaker BA, Simonyan V, Song JS, Thiessen PA, Yamashita RA, Yin JJ, Zhang D, Bryant SH: CDD: a Conserved Domain Database for protein classification. Nucleic Acids Res 2005, 33(Database issue):D192-6.
  36. ClustalW [http://www.ebi.ac.uk/Tools/clustalw2/index.html\]
  37. BoxShade [http://www.ch.embnet.org/software/BOX\_form.html\]