Abstract
In the Drosophila embryo, the alphaPS2betaPS integrin heterodimer is localized tightly at the termini of the multinucleate muscles where they attach to the alphaPS1betaPS-containing epidermal tendon cells. Here we examine the basis for alphaPS2betaPS integrin subcellular localization. We show that the betaPS cytoplasmic tail is sufficient to direct the localization of a heterologous transmembrane protein, CD2, to the muscle termini in vivo. This localization does not occur via an association with structures set up by the endogenous betaPS integrins, since it can occur even in the absence of the betaPS protein. Furthermore, the subcellular localization of the alphaPS2betaPS integrin is not dependent on any other interactions between the muscles and the tendon cells. In embryos that lack the segmental tendon cells, due to a mutation removing the related segment polarity genes engrailed and invected, alphaPS2betaPS is still localized to the muscle termini even though the ventral longitudinal muscles are not attached to the epidermis, but instead are attached end to end. Thus the alphaPS2betaPS integrin can be localized by an intracellular mechanism within the muscles. Our results challenge the view that the transmission of signals from the extracellular environment via integrins is required for the organization of the cytoskeleton and the resultant cellular polarity.
Full Text
The Full Text of this article is available as a PDF (3.2 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Armand P., Knapp A. C., Hirsch A. J., Wieschaus E. F., Cole M. D. A novel basic helix-loop-helix protein is expressed in muscle attachment sites of the Drosophila epidermis. Mol Cell Biol. 1994 Jun;14(6):4145–4154. doi: 10.1128/mcb.14.6.4145. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bate M. The embryonic development of larval muscles in Drosophila. Development. 1990 Nov;110(3):791–804. doi: 10.1242/dev.110.3.791. [DOI] [PubMed] [Google Scholar]
- 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]
- Borkowski O. M., Brown N. H., Bate M. Anterior-posterior subdivision and the diversification of the mesoderm in Drosophila. Development. 1995 Dec;121(12):4183–4193. doi: 10.1242/dev.121.12.4183. [DOI] [PubMed] [Google Scholar]
- Brabant M. C., Brower D. L. PS2 integrin requirements in Drosophila embryo and wing morphogenesis. Dev Biol. 1993 May;157(1):49–59. doi: 10.1006/dbio.1993.1111. [DOI] [PubMed] [Google Scholar]
- 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]
- Briesewitz R., Kern A., Marcantonio E. E. Ligand-dependent and -independent integrin focal contact localization: the role of the alpha chain cytoplasmic domain. Mol Biol Cell. 1993 Jun;4(6):593–604. doi: 10.1091/mbc.4.6.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brower D. L., Bunch T. A., Mukai L., Adamson T. E., Wehrli M., Lam S., Friedlander E., Roote C. E., Zusman S. Nonequivalent requirements for PS1 and PS2 integrin at cell attachments in Drosophila: genetic analysis of the alpha PS1 integrin subunit. Development. 1995 May;121(5):1311–1320. doi: 10.1242/dev.121.5.1311. [DOI] [PubMed] [Google Scholar]
- Brower D. L., Wilcox M., Piovant M., Smith R. J., Reger L. A. Related cell-surface antigens expressed with positional specificity in Drosophila imaginal discs. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7485–7489. doi: 10.1073/pnas.81.23.7485. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown N. H. Integrins hold Drosophila together. Bioessays. 1993 Jun;15(6):383–390. doi: 10.1002/bies.950150604. [DOI] [PubMed] [Google Scholar]
- Brown N. H. Null mutations in the alpha PS2 and beta PS integrin subunit genes have distinct phenotypes. Development. 1994 May;120(5):1221–1231. doi: 10.1242/dev.120.5.1221. [DOI] [PubMed] [Google Scholar]
- Bunch T. A., Salatino R., Engelsgjerd M. C., Mukai L., West R. F., Brower D. L. Characterization of mutant alleles of myospheroid, the gene encoding the beta subunit of the Drosophila PS integrins. Genetics. 1992 Oct;132(2):519–528. doi: 10.1093/genetics/132.2.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burridge K., Fath K., Kelly T., Nuckolls G., Turner C. Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol. 1988;4:487–525. doi: 10.1146/annurev.cb.04.110188.002415. [DOI] [PubMed] [Google Scholar]
- Cheresh D. A., Spiro R. C. Biosynthetic and functional properties of an Arg-Gly-Asp-directed receptor involved in human melanoma cell attachment to vitronectin, fibrinogen, and von Willebrand factor. J Biol Chem. 1987 Dec 25;262(36):17703–17711. [PubMed] [Google Scholar]
- Clark E. A., Brugge J. S. Integrins and signal transduction pathways: the road taken. Science. 1995 Apr 14;268(5208):233–239. doi: 10.1126/science.7716514. [DOI] [PubMed] [Google Scholar]
- D'Souza S. E., Ginsberg M. H., Burke T. A., Plow E. F. The ligand binding site of the platelet integrin receptor GPIIb-IIIa is proximal to the second calcium binding domain of its alpha subunit. J Biol Chem. 1990 Feb 25;265(6):3440–3446. [PubMed] [Google Scholar]
- Dunin-Borkowski O. M., Brown N. H. Mammalian CD2 is an effective heterologous marker of the cell surface in Drosophila. Dev Biol. 1995 Apr;168(2):689–693. doi: 10.1006/dbio.1995.1115. [DOI] [PubMed] [Google Scholar]
- Fath K. R., Edgell C. J., Burridge K. The distribution of distinct integrins in focal contacts is determined by the substratum composition. J Cell Sci. 1989 Jan;92(Pt 1):67–75. doi: 10.1242/jcs.92.1.67. [DOI] [PubMed] [Google Scholar]
- 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]
- Frasch M. Induction of visceral and cardiac mesoderm by ectodermal Dpp in the early Drosophila embryo. Nature. 1995 Mar 30;374(6521):464–467. doi: 10.1038/374464a0. [DOI] [PubMed] [Google Scholar]
- Fässler R., Meyer M. Consequences of lack of beta 1 integrin gene expression in mice. Genes Dev. 1995 Aug 1;9(15):1896–1908. doi: 10.1101/gad.9.15.1896. [DOI] [PubMed] [Google Scholar]
- Geiger B., Salomon D., Takeichi M., Hynes R. O. A chimeric N-cadherin/beta 1-integrin receptor which localizes to both cell-cell and cell-matrix adhesions. J Cell Sci. 1992 Dec;103(Pt 4):943–951. doi: 10.1242/jcs.103.4.943. [DOI] [PubMed] [Google Scholar]
- Gettner S. N., Kenyon C., Reichardt L. F. Characterization of beta pat-3 heterodimers, a family of essential integrin receptors in C. elegans. J Cell Biol. 1995 May;129(4):1127–1141. doi: 10.1083/jcb.129.4.1127. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gotwals P. J., Fessler L. I., Wehrli M., Hynes R. O. Drosophila PS1 integrin is a laminin receptor and differs in ligand specificity from PS2. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11447–11451. doi: 10.1073/pnas.91.24.11447. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Greig S., Akam M. Homeotic genes autonomously specify one aspect of pattern in the Drosophila mesoderm. Nature. 1993 Apr 15;362(6421):630–632. doi: 10.1038/362630a0. [DOI] [PubMed] [Google Scholar]
- Grinblat Y., Zusman S., Yee G., Hynes R. O., Kafatos F. C. Functions of the cytoplasmic domain of the beta PS integrin subunit during Drosophila development. Development. 1994 Jan;120(1):91–102. doi: 10.1242/dev.120.1.91. [DOI] [PubMed] [Google Scholar]
- Henchcliffe C., García-Alonso L., Tang J., Goodman C. S. Genetic analysis of laminin A reveals diverse functions during morphogenesis in Drosophila. Development. 1993 Jun;118(2):325–337. doi: 10.1242/dev.118.2.325. [DOI] [PubMed] [Google Scholar]
- Hidalgo A. Three distinct roles for the engrailed gene in Drosophila wing development. Curr Biol. 1994 Dec 1;4(12):1087–1098. doi: 10.1016/s0960-9822(00)00247-5. [DOI] [PubMed] [Google Scholar]
- Hresko M. C., Williams B. D., Waterston R. H. Assembly of body wall muscle and muscle cell attachment structures in Caenorhabditis elegans. J Cell Biol. 1994 Feb;124(4):491–506. doi: 10.1083/jcb.124.4.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Kishimoto T. K., Hollander N., Roberts T. M., Anderson D. C., Springer T. A. Heterogeneous mutations in the beta subunit common to the LFA-1, Mac-1, and p150,95 glycoproteins cause leukocyte adhesion deficiency. Cell. 1987 Jul 17;50(2):193–202. doi: 10.1016/0092-8674(87)90215-7. [DOI] [PubMed] [Google Scholar]
- LaFlamme S. E., Akiyama S. K., Yamada K. M. Regulation of fibronectin receptor distribution. J Cell Biol. 1992 Apr;117(2):437–447. doi: 10.1083/jcb.117.2.437. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LaFlamme S. E., Thomas L. A., Yamada S. S., Yamada K. M. Single subunit chimeric integrins as mimics and inhibitors of endogenous integrin functions in receptor localization, cell spreading and migration, and matrix assembly. J Cell Biol. 1994 Sep;126(5):1287–1298. doi: 10.1083/jcb.126.5.1287. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lawrence P. A., Johnston P. On the role of the engrailed+ gene in the internal organs of Drosophila. EMBO J. 1984 Dec 1;3(12):2839–2844. doi: 10.1002/j.1460-2075.1984.tb02217.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Lub M., van Kooyk Y., Figdor C. G. Ins and outs of LFA-1. Immunol Today. 1995 Oct;16(10):479–483. doi: 10.1016/0167-5699(95)80031-x. [DOI] [PubMed] [Google Scholar]
- Miyamoto S., Akiyama S. K., Yamada K. M. Synergistic roles for receptor occupancy and aggregation in integrin transmembrane function. Science. 1995 Feb 10;267(5199):883–885. doi: 10.1126/science.7846531. [DOI] [PubMed] [Google Scholar]
- Newman S. M., Jr, Wright T. R. A histological and ultrastructural analysis of developmental defects produced by the mutation, lethal(1)myospheroid, in Drosophila melanogaster. Dev Biol. 1981 Sep;86(2):393–402. doi: 10.1016/0012-1606(81)90197-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Rushton E., Drysdale R., Abmayr S. M., Michelson A. M., Bate M. Mutations in a novel gene, myoblast city, provide evidence in support of the founder cell hypothesis for Drosophila muscle development. Development. 1995 Jul;121(7):1979–1988. doi: 10.1242/dev.121.7.1979. [DOI] [PubMed] [Google Scholar]
- Smith J. W., Cheresh D. A. Integrin (alpha v beta 3)-ligand interaction. Identification of a heterodimeric RGD binding site on the vitronectin receptor. J Biol Chem. 1990 Feb 5;265(4):2168–2172. [PubMed] [Google Scholar]
- Stephens L. E., Sutherland A. E., Klimanskaya I. V., Andrieux A., Meneses J., Pedersen R. A., Damsky C. H. Deletion of beta 1 integrins in mice results in inner cell mass failure and peri-implantation lethality. Genes Dev. 1995 Aug 1;9(15):1883–1895. doi: 10.1101/gad.9.15.1883. [DOI] [PubMed] [Google Scholar]
- Turner C. E. Paxillin: a cytoskeletal target for tyrosine kinases. Bioessays. 1994 Jan;16(1):47–52. doi: 10.1002/bies.950160107. [DOI] [PubMed] [Google Scholar]
- 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]
- Volk T., VijayRaghavan K. A central role for epidermal segment border cells in the induction of muscle patterning in the Drosophila embryo. Development. 1994 Jan;120(1):59–70. doi: 10.1242/dev.120.1.59. [DOI] [PubMed] [Google Scholar]
- WRIGHT T. R. The phenogenetics of the embryonic mutant, lethal myospheroid, in Drosophila melanogaster. J Exp Zool. 1960 Feb;143:77–99. doi: 10.1002/jez.1401430107. [DOI] [PubMed] [Google Scholar]
- Williams A. F., Barclay A. N., Clark S. J., Paterson D. J., Willis A. C. Similarities in sequences and cellular expression between rat CD2 and CD4 antigens. J Exp Med. 1987 Feb 1;165(2):368–380. doi: 10.1084/jem.165.2.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Williams B. D., Waterston R. H. Genes critical for muscle development and function in Caenorhabditis elegans identified through lethal mutations. J Cell Biol. 1994 Feb;124(4):475–490. doi: 10.1083/jcb.124.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yang J. T., Rayburn H., Hynes R. O. Cell adhesion events mediated by alpha 4 integrins are essential in placental and cardiac development. Development. 1995 Feb;121(2):549–560. doi: 10.1242/dev.121.2.549. [DOI] [PubMed] [Google Scholar]
- Yang J. T., Rayburn H., Hynes R. O. Embryonic mesodermal defects in alpha 5 integrin-deficient mice. Development. 1993 Dec;119(4):1093–1105. doi: 10.1242/dev.119.4.1093. [DOI] [PubMed] [Google Scholar]
- Yarnitzky T., Volk T. Laminin is required for heart, somatic muscles, and gut development in the Drosophila embryo. Dev Biol. 1995 Jun;169(2):609–618. doi: 10.1006/dbio.1995.1173. [DOI] [PubMed] [Google Scholar]
- Ylänne J., Chen Y., O'Toole T. E., Loftus J. C., Takada Y., Ginsberg M. H. Distinct functions of integrin alpha and beta subunit cytoplasmic domains in cell spreading and formation of focal adhesions. J Cell Biol. 1993 Jul;122(1):223–233. doi: 10.1083/jcb.122.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]