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. 1996 Sep 2;134(6):1469–1482. doi: 10.1083/jcb.134.6.1469

Dlg protein is required for junction structure, cell polarity, and proliferation control in Drosophila epithelia

PMCID: PMC2120992  PMID: 8830775

Abstract

The Discs large (Dlg) protein of Drosophila is the prototypic member of a growing family of proteins termed membrane-associated guanylate kinase homologs (MAGUKs). The MAGUKs are composed of a series of peptide domains that include one or three DHR/PDZs, an SH3, and a region homologous to guanylate kinase (GUK). We have previously shown that the product of this gene, the Dlg protein, is localized at the septate junctions between epithelial cells, and that mutations in the gene cause neoplastic overgrowth of the imaginal discs. The dlg locus is therefore defined as a tumor suppressor gene. In this paper, we show that the Dlg protein is localized on the cytoplasmic face of the septate junction and is required for the maintenance of this structure. It is also required for proper organization of the cytoskeleton, for the differential localization of membrane proteins, and for apicobasal polarity of epithelial cells. However, these other functions can be uncoupled from Dlg's role as a tumor suppressor since mutations in two domains of the protein, the SH3 and GUK, cause loss of normal cell proliferation control without affecting the other functions of the protein. These results suggest that, besides regulating cellular proliferation, the Dlg protein is a critical component of the septate junctions and is required for maintaining apicobasal polarity in Drosophila epithelium.

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Selected References

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  1. Abbott L. A., Natzle J. E. Epithelial polarity and cell separation in the neoplastic l(1)dlg-1 mutant of Drosophila. Mech Dev. 1992 Mar;37(1-2):43–56. doi: 10.1016/0925-4773(92)90014-b. [DOI] [PubMed] [Google Scholar]
  2. Anderson J. M., Balda M. S., Fanning A. S. The structure and regulation of tight junctions. Curr Opin Cell Biol. 1993 Oct;5(5):772–778. doi: 10.1016/0955-0674(93)90024-k. [DOI] [PubMed] [Google Scholar]
  3. Balda M. S., Gonzalez-Mariscal L., Matter K., Cereijido M., Anderson J. M. Assembly of the tight junction: the role of diacylglycerol. J Cell Biol. 1993 Oct;123(2):293–302. doi: 10.1083/jcb.123.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bieber A. J., Snow P. M., Hortsch M., Patel N. H., Jacobs J. R., Traquina Z. R., Schilling J., Goodman C. S. Drosophila neuroglian: a member of the immunoglobulin superfamily with extensive homology to the vertebrate neural adhesion molecule L1. Cell. 1989 Nov 3;59(3):447–460. doi: 10.1016/0092-8674(89)90029-9. [DOI] [PubMed] [Google Scholar]
  5. Boedigheimer M., Laughon A. Expanded: a gene involved in the control of cell proliferation in imaginal discs. Development. 1993 Aug;118(4):1291–1301. doi: 10.1242/dev.118.4.1291. [DOI] [PubMed] [Google Scholar]
  6. Brand A. H., Perrimon N. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development. 1993 Jun;118(2):401–415. doi: 10.1242/dev.118.2.401. [DOI] [PubMed] [Google Scholar]
  7. Brenman J. E., Chao D. S., Gee S. H., McGee A. W., Craven S. E., Santillano D. R., Wu Z., Huang F., Xia H., Peters M. F. Interaction of nitric oxide synthase with the postsynaptic density protein PSD-95 and alpha1-syntrophin mediated by PDZ domains. Cell. 1996 Mar 8;84(5):757–767. doi: 10.1016/s0092-8674(00)81053-3. [DOI] [PubMed] [Google Scholar]
  8. Brower D. L., Smith R. J., Wilcox M. A monoclonal antibody specific for diploid epithelial cells in Drosophila. Nature. 1980 Jun 5;285(5764):403–405. doi: 10.1038/285403a0. [DOI] [PubMed] [Google Scholar]
  9. Bryant P. J., Watson K. L., Justice R. W., Woods D. F. Tumor suppressor genes encoding proteins required for cell interactions and signal transduction in Drosophila. Dev Suppl. 1993:239–249. [PubMed] [Google Scholar]
  10. Cereijido M., Contreras R. G., Gonzalez-Mariscal L. Development and alteration of polarity. Annu Rev Physiol. 1989;51:785–795. doi: 10.1146/annurev.ph.51.030189.004033. [DOI] [PubMed] [Google Scholar]
  11. Cho K. O., Hunt C. A., Kennedy M. B. The rat brain postsynaptic density fraction contains a homolog of the Drosophila discs-large tumor suppressor protein. Neuron. 1992 Nov;9(5):929–942. doi: 10.1016/0896-6273(92)90245-9. [DOI] [PubMed] [Google Scholar]
  12. Colombo A., Bonfanti P., Camatini M. Actin, alpha-actinin, and vinculin are associated with septate junctions in Insecta. Cell Motil Cytoskeleton. 1993;26(3):205–213. doi: 10.1002/cm.970260304. [DOI] [PubMed] [Google Scholar]
  13. Contreras R. G., Miller J. H., Zamora M., González-Mariscal L., Cereijido M. Interaction of calcium with plasma membrane of epithelial (MDCK) cells during junction formation. Am J Physiol. 1992 Aug;263(2 Pt 1):C313–C318. doi: 10.1152/ajpcell.1992.263.2.C313. [DOI] [PubMed] [Google Scholar]
  14. Duclos F., Boschert U., Sirugo G., Mandel J. L., Hen R., Koenig M. Gene in the region of the Friedreich ataxia locus encodes a putative transmembrane protein expressed in the nervous system. Proc Natl Acad Sci U S A. 1993 Jan 1;90(1):109–113. doi: 10.1073/pnas.90.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Eaton S., Simons K. Apical, basal, and lateral cues for epithelial polarization. Cell. 1995 Jul 14;82(1):5–8. doi: 10.1016/0092-8674(95)90045-4. [DOI] [PubMed] [Google Scholar]
  16. Fehon R. G., Dawson I. A., Artavanis-Tsakonas S. A Drosophila homologue of membrane-skeleton protein 4.1 is associated with septate junctions and is encoded by the coracle gene. Development. 1994 Mar;120(3):545–557. doi: 10.1242/dev.120.3.545. [DOI] [PubMed] [Google Scholar]
  17. Gonzalez-Mariscal L., Contreras R. G., Bolívar J. J., Ponce A., Chávez De Ramirez B., Cereijido M. Role of calcium in tight junction formation between epithelial cells. Am J Physiol. 1990 Dec;259(6 Pt 1):C978–C986. doi: 10.1152/ajpcell.1990.259.6.C978. [DOI] [PubMed] [Google Scholar]
  18. Gumbiner B. Generation and maintenance of epithelial cell polarity. Curr Opin Cell Biol. 1990 Oct;2(5):881–887. doi: 10.1016/0955-0674(90)90087-u. [DOI] [PubMed] [Google Scholar]
  19. Hoskins R., Hajnal A. F., Harp S. A., Kim S. K. The C. elegans vulval induction gene lin-2 encodes a member of the MAGUK family of cell junction proteins. Development. 1996 Jan;122(1):97–111. doi: 10.1242/dev.122.1.97. [DOI] [PubMed] [Google Scholar]
  20. Jesaitis L. A., Goodenough D. A. Molecular characterization and tissue distribution of ZO-2, a tight junction protein homologous to ZO-1 and the Drosophila discs-large tumor suppressor protein. J Cell Biol. 1994 Mar;124(6):949–961. doi: 10.1083/jcb.124.6.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kim S. K. Tight junctions, membrane-associated guanylate kinases and cell signaling. Curr Opin Cell Biol. 1995 Oct;7(5):641–649. doi: 10.1016/0955-0674(95)80105-7. [DOI] [PubMed] [Google Scholar]
  22. Kistner U., Garner C. C., Linial M. Nucleotide binding by the synapse associated protein SAP90. FEBS Lett. 1995 Feb 13;359(2-3):159–163. doi: 10.1016/0014-5793(95)00030-d. [DOI] [PubMed] [Google Scholar]
  23. Kistner U., Wenzel B. M., Veh R. W., Cases-Langhoff C., Garner A. M., Appeltauer U., Voss B., Gundelfinger E. D., Garner C. C. SAP90, a rat presynaptic protein related to the product of the Drosophila tumor suppressor gene dlg-A. J Biol Chem. 1993 Mar 5;268(7):4580–4583. [PubMed] [Google Scholar]
  24. Knust E., Tepass U., Wodarz A. crumbs and stardust, two genes of Drosophila required for the development of epithelial cell polarity. Dev Suppl. 1993:261–268. [PubMed] [Google Scholar]
  25. Kornau H. C., Schenker L. T., Kennedy M. B., Seeburg P. H. Domain interaction between NMDA receptor subunits and the postsynaptic density protein PSD-95. Science. 1995 Sep 22;269(5231):1737–1740. doi: 10.1126/science.7569905. [DOI] [PubMed] [Google Scholar]
  26. Lahey T., Gorczyca M., Jia X. X., Budnik V. The Drosophila tumor suppressor gene dlg is required for normal synaptic bouton structure. Neuron. 1994 Oct;13(4):823–835. doi: 10.1016/0896-6273(94)90249-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lue R. A., Marfatia S. M., Branton D., Chishti A. H. Cloning and characterization of hdlg: the human homologue of the Drosophila discs large tumor suppressor binds to protein 4.1. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9818–9822. doi: 10.1073/pnas.91.21.9818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Marfatia S. M., Lue R. A., Branton D., Chishti A. H. In vitro binding studies suggest a membrane-associated complex between erythroid p55, protein 4.1, and glycophorin C. J Biol Chem. 1994 Mar 25;269(12):8631–8634. [PubMed] [Google Scholar]
  29. Mazoyer S., Gayther S. A., Nagai M. A., Smith S. A., Dunning A., van Rensburg E. J., Albertsen H., White R., Ponder B. A. A gene (DLG2) located at 17q12-q21 encodes a new homologue of the Drosophila tumor suppressor dIg-A. Genomics. 1995 Jul 1;28(1):25–31. doi: 10.1006/geno.1995.1101. [DOI] [PubMed] [Google Scholar]
  30. McCormick F., Martin G. A., Clark R., Bollag G., Polakis P. Regulation of ras p21 by GTPase activating proteins. Cold Spring Harb Symp Quant Biol. 1991;56:237–241. doi: 10.1101/sqb.1991.056.01.029. [DOI] [PubMed] [Google Scholar]
  31. Müller B. M., Kistner U., Veh R. W., Cases-Langhoff C., Becker B., Gundelfinger E. D., Garner C. C. Molecular characterization and spatial distribution of SAP97, a novel presynaptic protein homologous to SAP90 and the Drosophila discs-large tumor suppressor protein. J Neurosci. 1995 Mar;15(3 Pt 2):2354–2366. doi: 10.1523/JNEUROSCI.15-03-02354.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Patel N. H., Snow P. M., Goodman C. S. Characterization and cloning of fasciclin III: a glycoprotein expressed on a subset of neurons and axon pathways in Drosophila. Cell. 1987 Mar 27;48(6):975–988. doi: 10.1016/0092-8674(87)90706-9. [DOI] [PubMed] [Google Scholar]
  33. Pawson T. SH2 and SH3 domains in signal transduction. Adv Cancer Res. 1994;64:87–110. doi: 10.1016/s0065-230x(08)60835-0. [DOI] [PubMed] [Google Scholar]
  34. Perrimon N. The maternal effect of lethal(1)discs-large-1: a recessive oncogene of Drosophila melanogaster. Dev Biol. 1988 Jun;127(2):392–407. doi: 10.1016/0012-1606(88)90326-0. [DOI] [PubMed] [Google Scholar]
  35. Ponting C. P., Phillips C. DHR domains in syntrophins, neuronal NO synthases and other intracellular proteins. Trends Biochem Sci. 1995 Mar;20(3):102–103. doi: 10.1016/s0968-0004(00)88973-2. [DOI] [PubMed] [Google Scholar]
  36. Ren R., Mayer B. J., Cicchetti P., Baltimore D. Identification of a ten-amino acid proline-rich SH3 binding site. Science. 1993 Feb 19;259(5098):1157–1161. doi: 10.1126/science.8438166. [DOI] [PubMed] [Google Scholar]
  37. Rhyu M. S., Jan L. Y., Jan Y. N. Asymmetric distribution of numb protein during division of the sensory organ precursor cell confers distinct fates to daughter cells. Cell. 1994 Feb 11;76(3):477–491. doi: 10.1016/0092-8674(94)90112-0. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Ruff P., Speicher D. W., Husain-Chishti A. Molecular identification of a major palmitoylated erythrocyte membrane protein containing the src homology 3 motif. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6595–6599. doi: 10.1073/pnas.88.15.6595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sato T., Irie S., Kitada S., Reed J. C. FAP-1: a protein tyrosine phosphatase that associates with Fas. Science. 1995 Apr 21;268(5209):411–415. doi: 10.1126/science.7536343. [DOI] [PubMed] [Google Scholar]
  41. Stehle T., Schulz G. E. Refined structure of the complex between guanylate kinase and its substrate GMP at 2.0 A resolution. J Mol Biol. 1992 Apr 20;224(4):1127–1141. doi: 10.1016/0022-2836(92)90474-x. [DOI] [PubMed] [Google Scholar]
  42. Stewart M., Murphy C., Fristrom J. W. The recovery and preliminary characterization of X chromosome mutants affecting imaginal discs of Drosophila melanogaster. Dev Biol. 1972 Jan;27(1):71–83. doi: 10.1016/0012-1606(72)90113-3. [DOI] [PubMed] [Google Scholar]
  43. Strand D., Jakobs R., Merdes G., Neumann B., Kalmes A., Heid H. W., Husmann I., Mechler B. M. The Drosophila lethal(2)giant larvae tumor suppressor protein forms homo-oligomers and is associated with nonmuscle myosin II heavy chain. J Cell Biol. 1994 Dec;127(5):1361–1373. doi: 10.1083/jcb.127.5.1361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Strand D., Raska I., Mechler B. M. The Drosophila lethal(2)giant larvae tumor suppressor protein is a component of the cytoskeleton. J Cell Biol. 1994 Dec;127(5):1345–1360. doi: 10.1083/jcb.127.5.1345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tepass U., Hartenstein V. The development of cellular junctions in the Drosophila embryo. Dev Biol. 1994 Feb;161(2):563–596. doi: 10.1006/dbio.1994.1054. [DOI] [PubMed] [Google Scholar]
  46. Tepass U., Knust E. Crumbs and stardust act in a genetic pathway that controls the organization of epithelia in Drosophila melanogaster. Dev Biol. 1993 Sep;159(1):311–326. doi: 10.1006/dbio.1993.1243. [DOI] [PubMed] [Google Scholar]
  47. Watson K. L., Konrad K. D., Woods D. F., Bryant P. J. Drosophila homolog of the human S6 ribosomal protein is required for tumor suppression in the hematopoietic system. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11302–11306. doi: 10.1073/pnas.89.23.11302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Willott E., Balda M. S., Fanning A. S., Jameson B., Van Itallie C., Anderson J. M. The tight junction protein ZO-1 is homologous to the Drosophila discs-large tumor suppressor protein of septate junctions. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7834–7838. doi: 10.1073/pnas.90.16.7834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wodarz A., Hinz U., Engelbert M., Knust E. Expression of crumbs confers apical character on plasma membrane domains of ectodermal epithelia of Drosophila. Cell. 1995 Jul 14;82(1):67–76. doi: 10.1016/0092-8674(95)90053-5. [DOI] [PubMed] [Google Scholar]
  50. Woods D. F., Bryant P. J. Apical junctions and cell signalling in epithelia. J Cell Sci Suppl. 1993;17:171–181. doi: 10.1242/jcs.1993.supplement_17.25. [DOI] [PubMed] [Google Scholar]
  51. Woods D. F., Bryant P. J. Molecular cloning of the lethal(1)discs large-1 oncogene of Drosophila. Dev Biol. 1989 Jul;134(1):222–235. doi: 10.1016/0012-1606(89)90092-4. [DOI] [PubMed] [Google Scholar]
  52. Woods D. F., Bryant P. J. The discs-large tumor suppressor gene of Drosophila encodes a guanylate kinase homolog localized at septate junctions. Cell. 1991 Aug 9;66(3):451–464. doi: 10.1016/0092-8674(81)90009-x. [DOI] [PubMed] [Google Scholar]
  53. Woods D. F., Bryant P. J. Tumor suppressor genes and signal transduction in Drosophila. Princess Takamatsu Symp. 1994;24:1–13. [PubMed] [Google Scholar]
  54. Woods D. F., Bryant P. J. ZO-1, DlgA and PSD-95/SAP90: homologous proteins in tight, septate and synaptic cell junctions. Mech Dev. 1993 Dec;44(2-3):85–89. doi: 10.1016/0925-4773(93)90059-7. [DOI] [PubMed] [Google Scholar]
  55. Zahraoui A., Joberty G., Arpin M., Fontaine J. J., Hellio R., Tavitian A., Louvard D. A small rab GTPase is distributed in cytoplasmic vesicles in non polarized cells but colocalizes with the tight junction marker ZO-1 in polarized epithelial cells. J Cell Biol. 1994 Jan;124(1-2):101–115. doi: 10.1083/jcb.124.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]

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