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. 1996 May;16(5):1896–1908. doi: 10.1128/mcb.16.5.1896

A Drosophila homolog of the Rac- and Cdc42-activated serine/threonine kinase PAK is a potential focal adhesion and focal complex protein that colocalizes with dynamic actin structures.

N Harden 1, J Lee 1, H Y Loh 1, Y M Ong 1, I Tan 1, T Leung 1, E Manser 1, L Lim 1
PMCID: PMC231177  PMID: 8628256

Abstract

Changes in cell morphology are essential in the development of a multicellular organism. The regulation of the cytoskeleton by the Rho subfamily of small GTP-binding proteins is an important determinant of cell shape. The Rho subfamily has been shown to participate in a variety of morphogenetic processes during Drosophila melanogaster development. We describe here a Drosophila homolog, DPAK, of the serine/threonine kinase PAK, a protein which is a target of the Rho subfamily proteins Rac and Cdc42. Rac, Cdc42, and PAK have previously been implicated in signaling by c-Jun amino-terminal kinases. DPAK bound to activated (GTP-bound) Drosophila Rac (DRacA) and Drosophila Cdc42. Similarities in the distributions of DPAK, integrin, and phosphotyrosine suggested an association of DPAK with focal adhesions and Cdc42- and Rac-induced focal adhesion-like focal complexes. DPAK was elevated in the leading edge of epidermal cells, whose morphological changes drive dorsal closure of the embryo. We have previously shown that the accumulation of cytoskeletal elements initiating cell shape changes in these cells could be inhibited by expression of a dominant-negative DRacA transgene. We show that leading-edge epidermal cells flanking segment borders, which express particularly large amounts of DPAK, undergo transient losses of cytoskeletal structures during dorsal closure. We propose that DPAK may be regulating the cytoskeleton through its association with focal adhesions and focal complexes and may be participating with DRacA in a c-Jun amino-terminal kinase signaling pathway recently demonstrated to be required for dorsal closure.

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Selected References

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  1. Adams A. E., Johnson D. I., Longnecker R. M., Sloat B. F., Pringle J. R. CDC42 and CDC43, two additional genes involved in budding and the establishment of cell polarity in the yeast Saccharomyces cerevisiae. J Cell Biol. 1990 Jul;111(1):131–142. doi: 10.1083/jcb.111.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bogaert T., Brown N., Wilcox M. The Drosophila PS2 antigen is an invertebrate integrin that, like the fibronectin receptor, becomes localized to muscle attachments. Cell. 1987 Dec 24;51(6):929–940. doi: 10.1016/0092-8674(87)90580-0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Brown N. H. Integrins hold Drosophila together. Bioessays. 1993 Jun;15(6):383–390. doi: 10.1002/bies.950150604. [DOI] [PubMed] [Google Scholar]
  5. Brown N. H., Kafatos F. C. Functional cDNA libraries from Drosophila embryos. J Mol Biol. 1988 Sep 20;203(2):425–437. doi: 10.1016/0022-2836(88)90010-1. [DOI] [PubMed] [Google Scholar]
  6. Chen W., Blanc J., Lim L. Characterization of a promiscuous GTPase-activating protein that has a Bcr-related domain from Caenorhabditis elegans. J Biol Chem. 1994 Jan 14;269(2):820–823. [PubMed] [Google Scholar]
  7. Chien C. B., Rosenthal D. E., Harris W. A., Holt C. E. Navigational errors made by growth cones without filopodia in the embryonic Xenopus brain. Neuron. 1993 Aug;11(2):237–251. doi: 10.1016/0896-6273(93)90181-p. [DOI] [PubMed] [Google Scholar]
  8. Coso O. A., Chiariello M., Yu J. C., Teramoto H., Crespo P., Xu N., Miki T., Gutkind J. S. The small GTP-binding proteins Rac1 and Cdc42 regulate the activity of the JNK/SAPK signaling pathway. Cell. 1995 Jun 30;81(7):1137–1146. doi: 10.1016/s0092-8674(05)80018-2. [DOI] [PubMed] [Google Scholar]
  9. Cvrcková F., De Virgilio C., Manser E., Pringle J. R., Nasmyth K. Ste20-like protein kinases are required for normal localization of cell growth and for cytokinesis in budding yeast. Genes Dev. 1995 Aug 1;9(15):1817–1830. doi: 10.1101/gad.9.15.1817. [DOI] [PubMed] [Google Scholar]
  10. Eaton S., Auvinen P., Luo L., Jan Y. N., Simons K. CDC42 and Rac1 control different actin-dependent processes in the Drosophila wing disc epithelium. J Cell Biol. 1995 Oct;131(1):151–164. doi: 10.1083/jcb.131.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fessler J. H., Fessler L. I. Drosophila extracellular matrix. Annu Rev Cell Biol. 1989;5:309–339. doi: 10.1146/annurev.cb.05.110189.001521. [DOI] [PubMed] [Google Scholar]
  12. Fogerty F. J., Fessler L. I., Bunch T. A., Yaron Y., Parker C. G., Nelson R. E., Brower D. L., Gullberg D., Fessler J. H. Tiggrin, a novel Drosophila extracellular matrix protein that functions as a ligand for Drosophila alpha PS2 beta PS integrins. Development. 1994 Jul;120(7):1747–1758. doi: 10.1242/dev.120.7.1747. [DOI] [PubMed] [Google Scholar]
  13. Glise B., Bourbon H., Noselli S. hemipterous encodes a novel Drosophila MAP kinase kinase, required for epithelial cell sheet movement. Cell. 1995 Nov 3;83(3):451–461. doi: 10.1016/0092-8674(95)90123-x. [DOI] [PubMed] [Google Scholar]
  14. Gotwals P. J., Paine-Saunders S. E., Stark K. A., Hynes R. O. Drosophila integrins and their ligands. Curr Opin Cell Biol. 1994 Oct;6(5):734–739. doi: 10.1016/0955-0674(94)90101-5. [DOI] [PubMed] [Google Scholar]
  15. Gout I., Dhand R., Hiles I. D., Fry M. J., Panayotou G., Das P., Truong O., Totty N. F., Hsuan J., Booker G. W. The GTPase dynamin binds to and is activated by a subset of SH3 domains. Cell. 1993 Oct 8;75(1):25–36. [PubMed] [Google Scholar]
  16. Gullberg D., Fessler L. I., Fessler J. H. Differentiation, extracellular matrix synthesis, and integrin assembly by Drosophila embryo cells cultured on vitronectin and laminin substrates. Dev Dyn. 1994 Feb;199(2):116–128. doi: 10.1002/aja.1001990205. [DOI] [PubMed] [Google Scholar]
  17. Hanks S. K., Calalb M. B., Harper M. C., Patel S. K. Focal adhesion protein-tyrosine kinase phosphorylated in response to cell attachment to fibronectin. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8487–8491. doi: 10.1073/pnas.89.18.8487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Harden N., Loh H. Y., Chia W., Lim L. A dominant inhibitory version of the small GTP-binding protein Rac disrupts cytoskeletal structures and inhibits developmental cell shape changes in Drosophila. Development. 1995 Mar;121(3):903–914. doi: 10.1242/dev.121.3.903. [DOI] [PubMed] [Google Scholar]
  19. Hariharan I. K., Hu K. Q., Asha H., Quintanilla A., Ezzell R. M., Settleman J. Characterization of rho GTPase family homologues in Drosophila melanogaster: overexpressing Rho1 in retinal cells causes a late developmental defect. EMBO J. 1995 Jan 16;14(2):292–302. doi: 10.1002/j.1460-2075.1995.tb07003.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Heidemann S. R., Lamoureux P., Buxbaum R. E. Growth cone behavior and production of traction force. J Cell Biol. 1990 Nov;111(5 Pt 1):1949–1957. doi: 10.1083/jcb.111.5.1949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Herskowitz I. MAP kinase pathways in yeast: for mating and more. Cell. 1995 Jan 27;80(2):187–197. doi: 10.1016/0092-8674(95)90402-6. [DOI] [PubMed] [Google Scholar]
  22. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  23. Ilić D., Furuta Y., Kanazawa S., Takeda N., Sobue K., Nakatsuji N., Nomura S., Fujimoto J., Okada M., Yamamoto T. Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice. Nature. 1995 Oct 12;377(6549):539–544. doi: 10.1038/377539a0. [DOI] [PubMed] [Google Scholar]
  24. Johnson D. I., Pringle J. R. Molecular characterization of CDC42, a Saccharomyces cerevisiae gene involved in the development of cell polarity. J Cell Biol. 1990 Jul;111(1):143–152. doi: 10.1083/jcb.111.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kiehart D. P., Feghali R. Cytoplasmic myosin from Drosophila melanogaster. J Cell Biol. 1986 Oct;103(4):1517–1525. doi: 10.1083/jcb.103.4.1517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kozma R., Ahmed S., Best A., Lim L. The Ras-related protein Cdc42Hs and bradykinin promote formation of peripheral actin microspikes and filopodia in Swiss 3T3 fibroblasts. Mol Cell Biol. 1995 Apr;15(4):1942–1952. doi: 10.1128/mcb.15.4.1942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Leberer E., Dignard D., Harcus D., Thomas D. Y., Whiteway M. The protein kinase homologue Ste20p is required to link the yeast pheromone response G-protein beta gamma subunits to downstream signalling components. EMBO J. 1992 Dec;11(13):4815–4824. doi: 10.1002/j.1460-2075.1992.tb05587.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Leptin M., Bogaert T., Lehmann R., Wilcox M. The function of PS integrins during Drosophila embryogenesis. Cell. 1989 Feb 10;56(3):401–408. doi: 10.1016/0092-8674(89)90243-2. [DOI] [PubMed] [Google Scholar]
  29. Luna E. J., Hitt A. L. Cytoskeleton--plasma membrane interactions. Science. 1992 Nov 6;258(5084):955–964. doi: 10.1126/science.1439807. [DOI] [PubMed] [Google Scholar]
  30. Luo L., Liao Y. J., Jan L. Y., Jan Y. N. Distinct morphogenetic functions of similar small GTPases: Drosophila Drac1 is involved in axonal outgrowth and myoblast fusion. Genes Dev. 1994 Aug 1;8(15):1787–1802. doi: 10.1101/gad.8.15.1787. [DOI] [PubMed] [Google Scholar]
  31. Maher P. A., Pasquale E. B., Wang J. Y., Singer S. J. Phosphotyrosine-containing proteins are concentrated in focal adhesions and intercellular junctions in normal cells. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6576–6580. doi: 10.1073/pnas.82.19.6576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Manser E., Leung T., Monfries C., Teo M., Hall C., Lim L. Diversity and versatility of GTPase activating proteins for the p21rho subfamily of ras G proteins detected by a novel overlay assay. J Biol Chem. 1992 Aug 15;267(23):16025–16028. [PubMed] [Google Scholar]
  33. Manser E., Leung T., Salihuddin H., Tan L., Lim L. A non-receptor tyrosine kinase that inhibits the GTPase activity of p21cdc42. Nature. 1993 May 27;363(6427):364–367. doi: 10.1038/363364a0. [DOI] [PubMed] [Google Scholar]
  34. Manser E., Leung T., Salihuddin H., Zhao Z. S., Lim L. A brain serine/threonine protein kinase activated by Cdc42 and Rac1. Nature. 1994 Jan 6;367(6458):40–46. doi: 10.1038/367040a0. [DOI] [PubMed] [Google Scholar]
  35. Marcus S., Polverino A., Chang E., Robbins D., Cobb M. H., Wigler M. H. Shk1, a homolog of the Saccharomyces cerevisiae Ste20 and mammalian p65PAK protein kinases, is a component of a Ras/Cdc42 signaling module in the fission yeast Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):6180–6184. doi: 10.1073/pnas.92.13.6180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Martin G. A., Bollag G., McCormick F., Abo A. A novel serine kinase activated by rac1/CDC42Hs-dependent autophosphorylation is related to PAK65 and STE20. EMBO J. 1995 May 1;14(9):1970–1978. doi: 10.1002/j.1460-2075.1995.tb07189.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Mayer B. J., Baltimore D. Signalling through SH2 and SH3 domains. Trends Cell Biol. 1993 Jan;3(1):8–13. doi: 10.1016/0962-8924(93)90194-6. [DOI] [PubMed] [Google Scholar]
  38. Minden A., Lin A., Claret F. X., Abo A., Karin M. Selective activation of the JNK signaling cascade and c-Jun transcriptional activity by the small GTPases Rac and Cdc42Hs. Cell. 1995 Jun 30;81(7):1147–1157. doi: 10.1016/s0092-8674(05)80019-4. [DOI] [PubMed] [Google Scholar]
  39. Nobes C. D., Hall A. Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell. 1995 Apr 7;81(1):53–62. doi: 10.1016/0092-8674(95)90370-4. [DOI] [PubMed] [Google Scholar]
  40. O'Connor T. P., Duerr J. S., Bentley D. Pioneer growth cone steering decisions mediated by single filopodial contacts in situ. J Neurosci. 1990 Dec;10(12):3935–3946. doi: 10.1523/JNEUROSCI.10-12-03935.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Olson M. F., Ashworth A., Hall A. An essential role for Rho, Rac, and Cdc42 GTPases in cell cycle progression through G1. Science. 1995 Sep 1;269(5228):1270–1272. doi: 10.1126/science.7652575. [DOI] [PubMed] [Google Scholar]
  42. Pesacreta T. C., Byers T. J., Dubreuil R., Kiehart D. P., Branton D. Drosophila spectrin: the membrane skeleton during embryogenesis. J Cell Biol. 1989 May;108(5):1697–1709. doi: 10.1083/jcb.108.5.1697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Polverino A., Frost J., Yang P., Hutchison M., Neiman A. M., Cobb M. H., Marcus S. Activation of mitogen-activated protein kinase cascades by p21-activated protein kinases in cell-free extracts of Xenopus oocytes. J Biol Chem. 1995 Nov 3;270(44):26067–26070. doi: 10.1074/jbc.270.44.26067. [DOI] [PubMed] [Google Scholar]
  44. Pombo C. M., Kehrl J. H., Sánchez I., Katz P., Avruch J., Zon L. I., Woodgett J. R., Force T., Kyriakis J. M. Activation of the SAPK pathway by the human STE20 homologue germinal centre kinase. Nature. 1995 Oct 26;377(6551):750–754. doi: 10.1038/377750a0. [DOI] [PubMed] [Google Scholar]
  45. Ramer S. W., Davis R. W. A dominant truncation allele identifies a gene, STE20, that encodes a putative protein kinase necessary for mating in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):452–456. doi: 10.1073/pnas.90.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. 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]
  47. Ridley A. J., Hall A. Signal transduction pathways regulating Rho-mediated stress fibre formation: requirement for a tyrosine kinase. EMBO J. 1994 Jun 1;13(11):2600–2610. doi: 10.1002/j.1460-2075.1994.tb06550.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Ridley A. J., Hall A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell. 1992 Aug 7;70(3):389–399. doi: 10.1016/0092-8674(92)90163-7. [DOI] [PubMed] [Google Scholar]
  49. Ridley A. J., Paterson H. F., Johnston C. L., Diekmann D., Hall A. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell. 1992 Aug 7;70(3):401–410. doi: 10.1016/0092-8674(92)90164-8. [DOI] [PubMed] [Google Scholar]
  50. Ring J. M., Martinez Arias A. puckered, a gene involved in position-specific cell differentiation in the dorsal epidermis of the Drosophila larva. Dev Suppl. 1993:251–259. [PubMed] [Google Scholar]
  51. Schaller M. D., Parsons J. T. Focal adhesion kinase and associated proteins. Curr Opin Cell Biol. 1994 Oct;6(5):705–710. doi: 10.1016/0955-0674(94)90097-3. [DOI] [PubMed] [Google Scholar]
  52. Schaller M. D., Parsons J. T. pp125FAK-dependent tyrosine phosphorylation of paxillin creates a high-affinity binding site for Crk. Mol Cell Biol. 1995 May;15(5):2635–2645. doi: 10.1128/mcb.15.5.2635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Shu S. Y., Ju G., Fan L. Z. The glucose oxidase-DAB-nickel method in peroxidase histochemistry of the nervous system. Neurosci Lett. 1988 Feb 29;85(2):169–171. doi: 10.1016/0304-3940(88)90346-1. [DOI] [PubMed] [Google Scholar]
  54. Simon M. N., De Virgilio C., Souza B., Pringle J. R., Abo A., Reed S. I. Role for the Rho-family GTPase Cdc42 in yeast mating-pheromone signal pathway. Nature. 1995 Aug 24;376(6542):702–705. doi: 10.1038/376702a0. [DOI] [PubMed] [Google Scholar]
  55. Sparks A. B., Quilliam L. A., Thorn J. M., Der C. J., Kay B. K. Identification and characterization of Src SH3 ligands from phage-displayed random peptide libraries. J Biol Chem. 1994 Sep 30;269(39):23853–23856. [PubMed] [Google Scholar]
  56. Turner C. E. Paxillin is a major phosphotyrosine-containing protein during embryonic development. J Cell Biol. 1991 Oct;115(1):201–207. doi: 10.1083/jcb.115.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Volk T., Fessler L. I., Fessler J. H. A role for integrin in the formation of sarcomeric cytoarchitecture. Cell. 1990 Nov 2;63(3):525–536. doi: 10.1016/0092-8674(90)90449-o. [DOI] [PubMed] [Google Scholar]
  58. Warn R. M., Magrath R., Webb S. Distribution of F-actin during cleavage of the Drosophila syncytial blastoderm. J Cell Biol. 1984 Jan;98(1):156–162. doi: 10.1083/jcb.98.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Young P. E., Richman A. M., Ketchum A. S., Kiehart D. P. Morphogenesis in Drosophila requires nonmuscle myosin heavy chain function. Genes Dev. 1993 Jan;7(1):29–41. doi: 10.1101/gad.7.1.29. [DOI] [PubMed] [Google Scholar]
  60. Zavortink M., Bunch T. A., Brower D. L. Functional properties of alternatively spliced forms of the Drosophila PS2 integrin alpha subunit. Cell Adhes Commun. 1993 Dec;1(3):251–264. doi: 10.3109/15419069309097258. [DOI] [PubMed] [Google Scholar]
  61. Zhang S., Han J., Sells M. A., Chernoff J., Knaus U. G., Ulevitch R. J., Bokoch G. M. Rho family GTPases regulate p38 mitogen-activated protein kinase through the downstream mediator Pak1. J Biol Chem. 1995 Oct 13;270(41):23934–23936. doi: 10.1074/jbc.270.41.23934. [DOI] [PubMed] [Google Scholar]
  62. Zhao Z. S., Leung T., Manser E., Lim L. Pheromone signalling in Saccharomyces cerevisiae requires the small GTP-binding protein Cdc42p and its activator CDC24. Mol Cell Biol. 1995 Oct;15(10):5246–5257. doi: 10.1128/mcb.15.10.5246. [DOI] [PMC free article] [PubMed] [Google Scholar]

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