Skip to main content
PMC home page
Genetics logoLink to Genetics
. 2003 Jun;164(2):553–563. doi: 10.1093/genetics/164.2.553

Distinct phospholipase C-gamma-dependent signaling pathways in the Drosophila eye and wing are revealed by a new small wing allele.

Rishikesh Mankidy 1, Jeremy Hastings 1, Justin R Thackeray 1
PMCID: PMC1462570  PMID: 12807776

Abstract

The Drosophila genome contains a single phospholipase C-gamma (PLC-gamma) homolog, encoded by small wing (sl), that acts as an inhibitor of receptor tyrosine kinase (RTK) signaling during photoreceptor R7 development. Although the existing sl alleles behave genetically as nulls, they may still produce truncated Sl products that could in theory still provide limited PLC-gamma function. Both to identify a true null allele and to probe structure-function relationships in Sl, we carried out an F(1) screen for new sl mutations and identified seven new alleles. Flies homozygous for any of these alleles are viable, with the same short-wing phenotype described previously; however, two of the alleles differ from any of those previously isolated in the severity of the eye phenotype: sl(9) homozygotes have a slightly more extreme extra-R7 phenotype, whereas sl(7) homozygotes have an almost wild-type eye. We determined the mutant defect in all seven alleles, revealing that sl(9) is a molecular null due to a very early stop codon, while sl(7) has a missense mutation in the highly conserved Y catalytic domain. Together with in vitro mutagenesis of the residue affected by the sl(7) mutation, these results confirm the role of Sl in RTK signaling and provide evidence for two genetically separable PLC-gamma-dependent pathways affecting the development of the eye and the wing.

Full Text

The Full Text of this article is available as a PDF (498.1 KB).

Selected References

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

  1. Adams M. D., Celniker S. E., Holt R. A., Evans C. A., Gocayne J. D., Amanatides P. G., Scherer S. E., Li P. W., Hoskins R. A., Galle R. F. The genome sequence of Drosophila melanogaster. Science. 2000 Mar 24;287(5461):2185–2195. doi: 10.1126/science.287.5461.2185. [DOI] [PubMed] [Google Scholar]
  2. Brennan C. A., Moses K. Determination of Drosophila photoreceptors: timing is everything. Cell Mol Life Sci. 2000 Feb;57(2):195–214. doi: 10.1007/PL00000684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chattopadhyay A., Vecchi M., Ji Q. s., Mernaugh R., Carpenter G. The role of individual SH2 domains in mediating association of phospholipase C-gamma1 with the activated EGF receptor. J Biol Chem. 1999 Sep 10;274(37):26091–26097. doi: 10.1074/jbc.274.37.26091. [DOI] [PubMed] [Google Scholar]
  4. Cheng H. F., Jiang M. J., Chen C. L., Liu S. M., Wong L. P., Lomasney J. W., King K. Cloning and identification of amino acid residues of human phospholipase C delta 1 essential for catalysis. J Biol Chem. 1995 Mar 10;270(10):5495–5505. doi: 10.1074/jbc.270.10.5495. [DOI] [PubMed] [Google Scholar]
  5. Diaz-Benjumea F. J., Hafen E. The sevenless signalling cassette mediates Drosophila EGF receptor function during epidermal development. Development. 1994 Mar;120(3):569–578. doi: 10.1242/dev.120.3.569. [DOI] [PubMed] [Google Scholar]
  6. Emori Y., Sugaya R., Akimaru H., Higashijima S., Shishido E., Saigo K., Homma Y. Drosophila phospholipase C-gamma expressed predominantly in blastoderm cells at cellularization and in endodermal cells during later embryonic stages. J Biol Chem. 1994 Jul 29;269(30):19474–19479. [PubMed] [Google Scholar]
  7. Essen L. O., Perisic O., Cheung R., Katan M., Williams R. L. Crystal structure of a mammalian phosphoinositide-specific phospholipase C delta. Nature. 1996 Apr 18;380(6575):595–602. doi: 10.1038/380595a0. [DOI] [PubMed] [Google Scholar]
  8. Falasca M., Logan S. K., Lehto V. P., Baccante G., Lemmon M. A., Schlessinger J. Activation of phospholipase C gamma by PI 3-kinase-induced PH domain-mediated membrane targeting. EMBO J. 1998 Jan 15;17(2):414–422. doi: 10.1093/emboj/17.2.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Freeman M. Cell determination strategies in the Drosophila eye. Development. 1997 Jan;124(2):261–270. doi: 10.1242/dev.124.2.261. [DOI] [PubMed] [Google Scholar]
  10. Gaul U., Mardon G., Rubin G. M. A putative Ras GTPase activating protein acts as a negative regulator of signaling by the Sevenless receptor tyrosine kinase. Cell. 1992 Mar 20;68(6):1007–1019. doi: 10.1016/0092-8674(92)90073-l. [DOI] [PubMed] [Google Scholar]
  11. Hashimoto A., Takeda K., Inaba M., Sekimata M., Kaisho T., Ikehara S., Homma Y., Akira S., Kurosaki T. Cutting edge: essential role of phospholipase C-gamma 2 in B cell development and function. J Immunol. 2000 Aug 15;165(4):1738–1742. doi: 10.4049/jimmunol.165.4.1738. [DOI] [PubMed] [Google Scholar]
  12. Hentze M. W., Kulozik A. E. A perfect message: RNA surveillance and nonsense-mediated decay. Cell. 1999 Feb 5;96(3):307–310. doi: 10.1016/s0092-8674(00)80542-5. [DOI] [PubMed] [Google Scholar]
  13. Huang J., Mohammadi M., Rodrigues G. A., Schlessinger J. Reduced activation of RAF-1 and MAP kinase by a fibroblast growth factor receptor mutant deficient in stimulation of phosphatidylinositol hydrolysis. J Biol Chem. 1995 Mar 10;270(10):5065–5072. doi: 10.1074/jbc.270.10.5065. [DOI] [PubMed] [Google Scholar]
  14. Huang P. S., Davis L., Huber H., Goodhart P. J., Wegrzyn R. E., Oliff A., Heimbrook D. C. An SH3 domain is required for the mitogenic activity of microinjected phospholipase C-gamma 1. FEBS Lett. 1995 Jan 30;358(3):287–292. doi: 10.1016/0014-5793(94)01453-8. [DOI] [PubMed] [Google Scholar]
  15. Ji Q. S., Winnier G. E., Niswender K. D., Horstman D., Wisdom R., Magnuson M. A., Carpenter G. Essential role of the tyrosine kinase substrate phospholipase C-gamma1 in mammalian growth and development. Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):2999–3003. doi: 10.1073/pnas.94.7.2999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Leading Group of the National Sample Survey of Fertility and Birth Control Communique of the National Sample Survey of Fertility and Birth Control (abstract). China Popul Newsl. 1989 Apr;6(2):8–10. [PubMed] [Google Scholar]
  17. Manning Charlene M., Mathews Wendy R., Fico Leah P., Thackeray Justin R. Phospholipase C-gamma contains introns shared by src homology 2 domains in many unrelated proteins. Genetics. 2003 Jun;164(2):433–442. doi: 10.1093/genetics/164.2.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Obermeier A., Tinhofer I., Grunicke H. H., Ullrich A. Transforming potentials of epidermal growth factor and nerve growth factor receptors inversely correlate with their phospholipase C gamma affinity and signal activation. EMBO J. 1996 Jan 2;15(1):73–82. [PMC free article] [PubMed] [Google Scholar]
  19. Orita M., Suzuki Y., Sekiya T., Hayashi K. Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics. 1989 Nov;5(4):874–879. doi: 10.1016/0888-7543(89)90129-8. [DOI] [PubMed] [Google Scholar]
  20. Powe A. C., Jr, Strathdee D., Cutforth T., D'Souza-Correia T., Gaines P., Thackeray J., Carlson J., Gaul U. In vivo functional analysis of Drosophila Gap1: involvement of Ca2+ and IP4 regulation. Mech Dev. 1999 Mar;81(1-2):89–101. doi: 10.1016/s0925-4773(98)00230-5. [DOI] [PubMed] [Google Scholar]
  21. Rameh L. E., Rhee S. G., Spokes K., Kazlauskas A., Cantley L. C., Cantley L. G. Phosphoinositide 3-kinase regulates phospholipase Cgamma-mediated calcium signaling. J Biol Chem. 1998 Sep 11;273(37):23750–23757. doi: 10.1074/jbc.273.37.23750. [DOI] [PubMed] [Google Scholar]
  22. Rebecchi M. J., Pentyala S. N. Structure, function, and control of phosphoinositide-specific phospholipase C. Physiol Rev. 2000 Oct;80(4):1291–1335. doi: 10.1152/physrev.2000.80.4.1291. [DOI] [PubMed] [Google Scholar]
  23. Rizo J., Südhof T. C. C2-domains, structure and function of a universal Ca2+-binding domain. J Biol Chem. 1998 Jun 26;273(26):15879–15882. doi: 10.1074/jbc.273.26.15879. [DOI] [PubMed] [Google Scholar]
  24. Ryu S. H., Suh P. G., Cho K. S., Lee K. Y., Rhee S. G. Bovine brain cytosol contains three immunologically distinct forms of inositolphospholipid-specific phospholipase C. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6649–6653. doi: 10.1073/pnas.84.19.6649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sivertzev-Dobzhansky N P, Dobzhansky T. Deficiency and Duplications for the Gene Bobbed in Drosophila Melanogaster. Genetics. 1933 May;18(3):173–192. doi: 10.1093/genetics/18.3.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tan P. B., Kim S. K. Signaling specificity: the RTK/RAS/MAP kinase pathway in metazoans. Trends Genet. 1999 Apr;15(4):145–149. doi: 10.1016/s0168-9525(99)01694-7. [DOI] [PubMed] [Google Scholar]
  27. Thackeray J. R., Gaines P. C., Ebert P., Carlson J. R. small wing encodes a phospholipase C-(gamma) that acts as a negative regulator of R7 development in Drosophila. Development. 1998 Dec;125(24):5033–5042. doi: 10.1242/dev.125.24.5033. [DOI] [PubMed] [Google Scholar]
  28. Wang D., Feng J., Wen R., Marine J. C., Sangster M. Y., Parganas E., Hoffmeyer A., Jackson C. W., Cleveland J. L., Murray P. J. Phospholipase Cgamma2 is essential in the functions of B cell and several Fc receptors. Immunity. 2000 Jul;13(1):25–35. doi: 10.1016/s1074-7613(00)00005-4. [DOI] [PubMed] [Google Scholar]
  29. Wang L. P., Lim C., Kuan Y., Chen C. L., Chen H. F., King K. Positive charge at position 549 is essential for phosphatidylinositol 4,5-bisphosphate-hydrolyzing but not phosphatidylinositol-hydrolyzing activities of human phospholipase C delta1. J Biol Chem. 1996 Oct 4;271(40):24505–24516. doi: 10.1074/jbc.271.40.24505. [DOI] [PubMed] [Google Scholar]
  30. Ye Keqiang, Aghdasi Bahman, Luo Hongbo R., Moriarity John L., Wu Frederick Y., Hong Jenny J., Hurt K. Joseph, Bae Sun Sik, Suh Pann-Ghill, Snyder Solomon H. Phospholipase C gamma 1 is a physiological guanine nucleotide exchange factor for the nuclear GTPase PIKE. Nature. 2002 Jan 31;415(6871):541–544. doi: 10.1038/415541a. [DOI] [PubMed] [Google Scholar]
  31. Zipursky S. L., Rubin G. M. Determination of neuronal cell fate: lessons from the R7 neuron of Drosophila. Annu Rev Neurosci. 1994;17:373–397. doi: 10.1146/annurev.ne.17.030194.002105. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES