Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1997 Dec;17(12):6859–6867. doi: 10.1128/mcb.17.12.6859

Transmembrane glycoprotein gp150 is a substrate for receptor tyrosine phosphatase DPTP10D in Drosophila cells.

S J Fashena 1, K Zinn 1
PMCID: PMC232542  PMID: 9372917

Abstract

We have begun to explore the downstream signaling pathways of receptor protein tyrosine phosphatases (RPTPs) that control axon guidance decisions in the Drosophila central nervous system. We have focused our studies on the adhesion molecule-like gp150 protein, which binds directly to and is an in vitro substrate for the RPTP DPTP10D. Here we show that gp150 and DPTP10D form stable complexes in Drosophila Schneider 2 (S2) cells and in wild-type larval tissue. We also demonstrate that the DPTP10D cytoplasmic domain is sufficient to confer binding to gp150. gp150 has a short cytoplasmic domain containing four tyrosines, all found within sequences similar to immunoreceptor family tyrosine-based activation motifs (ITAMs). We demonstrate that gp150 is tyrosine phosphorylated in wild-type larvae. In S2 cells, gp150 becomes tyrosine phosphorylated following incubation with PTP inhibitors or upon coexpression of the Dsrc tyrosine kinase. Phosphorylated Dsrc and an unknown 40-kDa phosphoprotein form stable complexes with gp150, thereby implicating them in a putative gp150 signaling pathway. When coexpressed with gp150, either full-length DPTP10D or its cytoplasmic domain mediates gp150 dephosphorylation whereas a catalytically inactive DPTP10D cytoplasmic domain does not. The neural RPTP DPTP99A can also induce gp150 dephosphorylation but does not coimmunoprecipitate with gp150. Taken together, the results suggest that gp150 transduces signals via phosphorylation of its ITAM-like elements. Phosphotyrosines on gp150 might function as binding sites for downstream signaling molecules, thereby initiating a signaling cascade that could be modulated in vivo by RPTPs such as DPTP10D.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Barnea G., Grumet M., Milev P., Silvennoinen O., Levy J. B., Sap J., Schlessinger J. Receptor tyrosine phosphatase beta is expressed in the form of proteoglycan and binds to the extracellular matrix protein tenascin. J Biol Chem. 1994 May 20;269(20):14349–14352. [PubMed] [Google Scholar]
  2. Bodden K., Bixby J. L. CRYP-2: a receptor-type tyrosine phosphatase selectively expressed by developing vertebrate neurons. J Neurobiol. 1996 Nov;31(3):309–324. doi: 10.1002/(SICI)1097-4695(199611)31:3<309::AID-NEU4>3.0.CO;2-E. [DOI] [PubMed] [Google Scholar]
  3. Brady-Kalnay S. M., Flint A. J., Tonks N. K. Homophilic binding of PTP mu, a receptor-type protein tyrosine phosphatase, can mediate cell-cell aggregation. J Cell Biol. 1993 Aug;122(4):961–972. doi: 10.1083/jcb.122.4.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bunch T. A., Grinblat Y., Goldstein L. S. Characterization and use of the Drosophila metallothionein promoter in cultured Drosophila melanogaster cells. Nucleic Acids Res. 1988 Feb 11;16(3):1043–1061. doi: 10.1093/nar/16.3.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cambier J. C. New nomenclature for the Reth motif (or ARH1/TAM/ARAM/YXXL) Immunol Today. 1995 Feb;16(2):110–110. doi: 10.1016/0167-5699(95)80105-7. [DOI] [PubMed] [Google Scholar]
  6. Desai C. J., Gindhart J. G., Jr, Goldstein L. S., Zinn K. Receptor tyrosine phosphatases are required for motor axon guidance in the Drosophila embryo. Cell. 1996 Feb 23;84(4):599–609. doi: 10.1016/s0092-8674(00)81035-1. [DOI] [PubMed] [Google Scholar]
  7. Desai C. J., Krueger N. X., Saito H., Zinn K. Competition and cooperation among receptor tyrosine phosphatases control motoneuron growth cone guidance in Drosophila. Development. 1997 May;124(10):1941–1952. doi: 10.1242/dev.124.10.1941. [DOI] [PubMed] [Google Scholar]
  8. Desai C. J., Popova E., Zinn K. A Drosophila receptor tyrosine phosphatase expressed in the embryonic CNS and larval optic lobes is a member of the set of proteins bearing the "HRP" carbohydrate epitope. J Neurosci. 1994 Dec;14(12):7272–7283. doi: 10.1523/JNEUROSCI.14-12-07272.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fang K. S., Sabe H., Saito H., Hanafusa H. Comparative study of three protein-tyrosine phosphatases. Chicken protein-tyrosine phosphatase lambda dephosphorylates c-Src tyrosine 527. J Biol Chem. 1994 Aug 5;269(31):20194–20200. [PubMed] [Google Scholar]
  10. Fantus I. G., Kadota S., Deragon G., Foster B., Posner B. I. Pervanadate [peroxide(s) of vanadate] mimics insulin action in rat adipocytes via activation of the insulin receptor tyrosine kinase. Biochemistry. 1989 Oct 31;28(22):8864–8871. doi: 10.1021/bi00448a027. [DOI] [PubMed] [Google Scholar]
  11. Fashena S. J., Zinn K. Cell-cell signaling: The ins and outs of receptor tyrosine phosphatases. Curr Biol. 1995 Dec 1;5(12):1367–1369. doi: 10.1016/s0960-9822(95)00272-7. [DOI] [PubMed] [Google Scholar]
  12. Flint A. J., Tiganis T., Barford D., Tonks N. K. Development of "substrate-trapping" mutants to identify physiological substrates of protein tyrosine phosphatases. Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):1680–1685. doi: 10.1073/pnas.94.5.1680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Furukawa T., Itoh M., Krueger N. X., Streuli M., Saito H. Specific interaction of the CD45 protein-tyrosine phosphatase with tyrosine-phosphorylated CD3 zeta chain. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10928–10932. doi: 10.1073/pnas.91.23.10928. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Garton A. J., Flint A. J., Tonks N. K. Identification of p130(cas) as a substrate for the cytosolic protein tyrosine phosphatase PTP-PEST. Mol Cell Biol. 1996 Nov;16(11):6408–6418. doi: 10.1128/mcb.16.11.6408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gertler F. B., Comer A. R., Juang J. L., Ahern S. M., Clark M. J., Liebl E. C., Hoffmann F. M. enabled, a dosage-sensitive suppressor of mutations in the Drosophila Abl tyrosine kinase, encodes an Abl substrate with SH3 domain-binding properties. Genes Dev. 1995 Mar 1;9(5):521–533. doi: 10.1101/gad.9.5.521. [DOI] [PubMed] [Google Scholar]
  16. Gertler F. B., Hill K. K., Clark M. J., Hoffmann F. M. Dosage-sensitive modifiers of Drosophila abl tyrosine kinase function: prospero, a regulator of axonal outgrowth, and disabled, a novel tyrosine kinase substrate. Genes Dev. 1993 Mar;7(3):441–453. doi: 10.1101/gad.7.3.441. [DOI] [PubMed] [Google Scholar]
  17. Hariharan I. K., Chuang P. T., Rubin G. M. Cloning and characterization of a receptor-class phosphotyrosine phosphatase gene expressed on central nervous system axons in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11266–11270. doi: 10.1073/pnas.88.24.11266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hashimoto C., Hudson K. L., Anderson K. V. The Toll gene of Drosophila, required for dorsal-ventral embryonic polarity, appears to encode a transmembrane protein. Cell. 1988 Jan 29;52(2):269–279. doi: 10.1016/0092-8674(88)90516-8. [DOI] [PubMed] [Google Scholar]
  19. Johnson S. A., Pleiman C. M., Pao L., Schneringer J., Hippen K., Cambier J. C. Phosphorylated immunoreceptor signaling motifs (ITAMs) exhibit unique abilities to bind and activate Lyn and Syk tyrosine kinases. J Immunol. 1995 Nov 15;155(10):4596–4603. [PubMed] [Google Scholar]
  20. Kavanaugh W. M., Williams L. T. An alternative to SH2 domains for binding tyrosine-phosphorylated proteins. Science. 1994 Dec 16;266(5192):1862–1865. doi: 10.1126/science.7527937. [DOI] [PubMed] [Google Scholar]
  21. Krueger N. X., Van Vactor D., Wan H. I., Gelbart W. M., Goodman C. S., Saito H. The transmembrane tyrosine phosphatase DLAR controls motor axon guidance in Drosophila. Cell. 1996 Feb 23;84(4):611–622. doi: 10.1016/s0092-8674(00)81036-3. [DOI] [PubMed] [Google Scholar]
  22. Kussick S. J., Cooper J. A. Overexpressed Drosophila src 64B is phosphorylated at its carboxy-terminal tyrosine, but is not catalytically repressed, in cultured Drosophila cells. Oncogene. 1992 Dec;7(12):2461–2470. [PubMed] [Google Scholar]
  23. Lai K. M., Olivier J. P., Gish G. D., Henkemeyer M., McGlade J., Pawson T. A Drosophila shc gene product is implicated in signaling by the DER receptor tyrosine kinase. Mol Cell Biol. 1995 Sep;15(9):4810–4818. doi: 10.1128/mcb.15.9.4810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lin D. M., Goodman C. S. Ectopic and increased expression of Fasciclin II alters motoneuron growth cone guidance. Neuron. 1994 Sep;13(3):507–523. doi: 10.1016/0896-6273(94)90022-1. [DOI] [PubMed] [Google Scholar]
  25. Mauro L. J., Dixon J. E. 'Zip codes' direct intracellular protein tyrosine phosphatases to the correct cellular 'address'. Trends Biochem Sci. 1994 Apr;19(4):151–155. doi: 10.1016/0968-0004(94)90274-7. [DOI] [PubMed] [Google Scholar]
  26. Milev P., Friedlander D. R., Sakurai T., Karthikeyan L., Flad M., Margolis R. K., Grumet M., Margolis R. U. Interactions of the chondroitin sulfate proteoglycan phosphacan, the extracellular domain of a receptor-type protein tyrosine phosphatase, with neurons, glia, and neural cell adhesion molecules. J Cell Biol. 1994 Dec;127(6 Pt 1):1703–1715. doi: 10.1083/jcb.127.6.1703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Neel B. G., Tonks N. K. Protein tyrosine phosphatases in signal transduction. Curr Opin Cell Biol. 1997 Apr;9(2):193–204. doi: 10.1016/s0955-0674(97)80063-4. [DOI] [PubMed] [Google Scholar]
  28. Nose A., Mahajan V. B., Goodman C. S. Connectin: a homophilic cell adhesion molecule expressed on a subset of muscles and the motoneurons that innervate them in Drosophila. Cell. 1992 Aug 21;70(4):553–567. doi: 10.1016/0092-8674(92)90426-d. [DOI] [PubMed] [Google Scholar]
  29. Olivier J. P., Raabe T., Henkemeyer M., Dickson B., Mbamalu G., Margolis B., Schlessinger J., Hafen E., Pawson T. A Drosophila SH2-SH3 adaptor protein implicated in coupling the sevenless tyrosine kinase to an activator of Ras guanine nucleotide exchange, Sos. Cell. 1993 Apr 9;73(1):179–191. doi: 10.1016/0092-8674(93)90170-u. [DOI] [PubMed] [Google Scholar]
  30. Peles E., Nativ M., Campbell P. L., Sakurai T., Martinez R., Lev S., Clary D. O., Schilling J., Barnea G., Plowman G. D. The carbonic anhydrase domain of receptor tyrosine phosphatase beta is a functional ligand for the axonal cell recognition molecule contactin. Cell. 1995 Jul 28;82(2):251–260. doi: 10.1016/0092-8674(95)90312-7. [DOI] [PubMed] [Google Scholar]
  31. Peles E., Nativ M., Lustig M., Grumet M., Schilling J., Martinez R., Plowman G. D., Schlessinger J. Identification of a novel contactin-associated transmembrane receptor with multiple domains implicated in protein-protein interactions. EMBO J. 1997 Mar 3;16(5):978–988. doi: 10.1093/emboj/16.5.978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Reinke R., Krantz D. E., Yen D., Zipursky S. L. Chaoptin, a cell surface glycoprotein required for Drosophila photoreceptor cell morphogenesis, contains a repeat motif found in yeast and human. Cell. 1988 Jan 29;52(2):291–301. doi: 10.1016/0092-8674(88)90518-1. [DOI] [PubMed] [Google Scholar]
  33. Resh M. D. Myristylation and palmitylation of Src family members: the fats of the matter. Cell. 1994 Feb 11;76(3):411–413. doi: 10.1016/0092-8674(94)90104-x. [DOI] [PubMed] [Google Scholar]
  34. Sap J., Jiang Y. P., Friedlander D., Grumet M., Schlessinger J. Receptor tyrosine phosphatase R-PTP-kappa mediates homophilic binding. Mol Cell Biol. 1994 Jan;14(1):1–9. doi: 10.1128/mcb.14.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schneider I. Cell lines derived from late embryonic stages of Drosophila melanogaster. J Embryol Exp Morphol. 1972 Apr;27(2):353–365. [PubMed] [Google Scholar]
  36. Simon M. A., Dodson G. S., Rubin G. M. An SH3-SH2-SH3 protein is required for p21Ras1 activation and binds to sevenless and Sos proteins in vitro. Cell. 1993 Apr 9;73(1):169–177. doi: 10.1016/0092-8674(93)90169-q. [DOI] [PubMed] [Google Scholar]
  37. Snow P. M., Bieber A. J., Goodman C. S. Fasciclin III: a novel homophilic adhesion molecule in Drosophila. Cell. 1989 Oct 20;59(2):313–323. doi: 10.1016/0092-8674(89)90293-6. [DOI] [PubMed] [Google Scholar]
  38. Songyang Z., Carraway K. L., 3rd, Eck M. J., Harrison S. C., Feldman R. A., Mohammadi M., Schlessinger J., Hubbard S. R., Smith D. P., Eng C. Catalytic specificity of protein-tyrosine kinases is critical for selective signalling. Nature. 1995 Feb 9;373(6514):536–539. doi: 10.1038/373536a0. [DOI] [PubMed] [Google Scholar]
  39. Songyang Z., Shoelson S. E., Chaudhuri M., Gish G., Pawson T., Haser W. G., King F., Roberts T., Ratnofsky S., Lechleider R. J. SH2 domains recognize specific phosphopeptide sequences. Cell. 1993 Mar 12;72(5):767–778. doi: 10.1016/0092-8674(93)90404-e. [DOI] [PubMed] [Google Scholar]
  40. Stoker A. W., Gehrig B., Haj F., Bay B. H. Axonal localisation of the CAM-like tyrosine phosphatase CRYP alpha: a signalling molecule of embryonic growth cones. Development. 1995 Jun;121(6):1833–1844. doi: 10.1242/dev.121.6.1833. [DOI] [PubMed] [Google Scholar]
  41. Stoker A. W., Gehrig B., Newton M. R., Bay B. H. Comparative localisation of CRYP alpha, a CAM-like tyrosine phosphatase, and NgCAM in the developing chick visual system. Brain Res Dev Brain Res. 1995 Dec 21;90(1-2):129–140. doi: 10.1016/0165-3806(96)83493-6. [DOI] [PubMed] [Google Scholar]
  42. Streuli M., Krueger N. X., Tsai A. Y., Saito H. A family of receptor-linked protein tyrosine phosphatases in humans and Drosophila. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8698–8702. doi: 10.1073/pnas.86.22.8698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tian S. S., Tsoulfas P., Zinn K. Three receptor-linked protein-tyrosine phosphatases are selectively expressed on central nervous system axons in the Drosophila embryo. Cell. 1991 Nov 15;67(4):675–685. doi: 10.1016/0092-8674(91)90063-5. [DOI] [PubMed] [Google Scholar]
  44. Tian S. S., Zinn K. An adhesion molecule-like protein that interacts with and is a substrate for a Drosophila receptor-linked protein tyrosine phosphatase. J Biol Chem. 1994 Nov 11;269(45):28478–28486. [PubMed] [Google Scholar]
  45. Trowbridge I. S., Thomas M. L. CD45: an emerging role as a protein tyrosine phosphatase required for lymphocyte activation and development. Annu Rev Immunol. 1994;12:85–116. doi: 10.1146/annurev.iy.12.040194.000505. [DOI] [PubMed] [Google Scholar]
  46. Yang X. H., Seow K. T., Bahri S. M., Oon S. H., Chia W. Two Drosophila receptor-like tyrosine phosphatase genes are expressed in a subset of developing axons and pioneer neurons in the embryonic CNS. Cell. 1991 Nov 15;67(4):661–673. doi: 10.1016/0092-8674(91)90062-4. [DOI] [PubMed] [Google Scholar]
  47. Zheng X. M., Wang Y., Pallen C. J. Cell transformation and activation of pp60c-src by overexpression of a protein tyrosine phosphatase. Nature. 1992 Sep 24;359(6393):336–339. doi: 10.1038/359336a0. [DOI] [PubMed] [Google Scholar]
  48. Zick Y., Sagi-Eisenberg R. A combination of H2O2 and vanadate concomitantly stimulates protein tyrosine phosphorylation and polyphosphoinositide breakdown in different cell lines. Biochemistry. 1990 Nov 6;29(44):10240–10245. doi: 10.1021/bi00496a013. [DOI] [PubMed] [Google Scholar]
  49. Zondag G. C., Koningstein G. M., Jiang Y. P., Sap J., Moolenaar W. H., Gebbink M. F. Homophilic interactions mediated by receptor tyrosine phosphatases mu and kappa. A critical role for the novel extracellular MAM domain. J Biol Chem. 1995 Jun 16;270(24):14247–14250. doi: 10.1074/jbc.270.24.14247. [DOI] [PubMed] [Google Scholar]
  50. den Hertog J., Pals C. E., Peppelenbosch M. P., Tertoolen L. G., de Laat S. W., Kruijer W. Receptor protein tyrosine phosphatase alpha activates pp60c-src and is involved in neuronal differentiation. EMBO J. 1993 Oct;12(10):3789–3798. doi: 10.1002/j.1460-2075.1993.tb06057.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES