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. 1994 May 1;125(3):607–616. doi: 10.1083/jcb.125.3.607

Identification of secreted and cytosolic gelsolin in Drosophila

PMCID: PMC2119988  PMID: 8175883

Abstract

We have cloned the gene for Drosophila gelsolin. Two mRNAs are produced from this gene by differential splicing. The protein encoded by the longer mRNA has a signal peptide and its electrophoretic mobility when translated in vitro in the presence of microsomes is higher than when it is translated without microsomes. The protein translated from the shorter mRNA does not show this difference. This indicates that Drosophila like vertebrates has two forms of gelsolin, one secreted, the other cytoplasmic. The mRNA for both is present ubiquitously in the early embryo. Later, the cytoplasmic form is expressed in parts of the gut. The RNA for the secreted form is expressed in the fat body, and the secreted protein is abundant in extracellular fluid (hemolymph). The cytoplasmic form of gelsolin co-localizes with F-actin in the cortex of the cells in the embryo and in larval epithelia. However, during cellularization of the blastoderm it is reduced at the base of the cleavage furrow, a structure similar to the contractile ring in dividing cells.

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Selected References

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  1. Ampe C., Vandekerckhove J. The F-actin capping proteins of Physarum polycephalum: cap42(a) is very similar, if not identical, to fragmin and is structurally and functionally very homologous to gelsolin; cap42(b) is Physarum actin. EMBO J. 1987 Dec 20;6(13):4149–4157. doi: 10.1002/j.1460-2075.1987.tb02761.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. André E., Brink M., Gerisch G., Isenberg G., Noegel A., Schleicher M., Segall J. E., Wallraff E. A Dictyostelium mutant deficient in severin, an F-actin fragmenting protein, shows normal motility and chemotaxis. J Cell Biol. 1989 Mar;108(3):985–995. doi: 10.1083/jcb.108.3.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. André E., Lottspeich F., Schleicher M., Noegel A. Severin, gelsolin, and villin share a homologous sequence in regions presumed to contain F-actin severing domains. J Biol Chem. 1988 Jan 15;263(2):722–727. [PubMed] [Google Scholar]
  4. Arpin M., Pringault E., Finidori J., Garcia A., Jeltsch J. M., Vandekerckhove J., Louvard D. Sequence of human villin: a large duplicated domain homologous with other actin-severing proteins and a unique small carboxy-terminal domain related to villin specificity. J Cell Biol. 1988 Nov;107(5):1759–1766. doi: 10.1083/jcb.107.5.1759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bazari W. L., Matsudaira P., Wallek M., Smeal T., Jakes R., Ahmed Y. Villin sequence and peptide map identify six homologous domains. Proc Natl Acad Sci U S A. 1988 Jul;85(14):4986–4990. doi: 10.1073/pnas.85.14.4986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Brown N. H., King D. L., Wilcox M., Kafatos F. C. Developmentally regulated alternative splicing of Drosophila integrin PS2 alpha transcripts. Cell. 1989 Oct 6;59(1):185–195. doi: 10.1016/0092-8674(89)90880-5. [DOI] [PubMed] [Google Scholar]
  8. Carron C. P., Hwo S. Y., Dingus J., Benson D. M., Meza I., Bryan J. A re-evaluation of cytoplasmic gelsolin localization. J Cell Biol. 1986 Jan;102(1):237–245. doi: 10.1083/jcb.102.1.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cunningham C. C., Stossel T. P., Kwiatkowski D. J. Enhanced motility in NIH 3T3 fibroblasts that overexpress gelsolin. Science. 1991 Mar 8;251(4998):1233–1236. doi: 10.1126/science.1848726. [DOI] [PubMed] [Google Scholar]
  10. Driever W., Siegel V., Nüsslein-Volhard C. Autonomous determination of anterior structures in the early Drosophila embryo by the bicoid morphogen. Development. 1990 Aug;109(4):811–820. doi: 10.1242/dev.109.4.811. [DOI] [PubMed] [Google Scholar]
  11. Dubreuil R. R., Byers T. J., Stewart C. T., Kiehart D. P. A beta-spectrin isoform from Drosophila (beta H) is similar in size to vertebrate dystrophin. J Cell Biol. 1990 Nov;111(5 Pt 1):1849–1858. doi: 10.1083/jcb.111.5.1849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dubreuil R., Byers T. J., Branton D., Goldstein L. S., Kiehart D. P. Drosophilia spectrin. I. Characterization of the purified protein. J Cell Biol. 1987 Nov;105(5):2095–2102. doi: 10.1083/jcb.105.5.2095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eichinger L., Noegel A. A., Schleicher M. Domain structure in actin-binding proteins: expression and functional characterization of truncated severin. J Cell Biol. 1991 Feb;112(4):665–676. doi: 10.1083/jcb.112.4.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Forscher P. Calcium and polyphosphoinositide control of cytoskeletal dynamics. Trends Neurosci. 1989 Nov;12(11):468–474. doi: 10.1016/0166-2236(89)90098-2. [DOI] [PubMed] [Google Scholar]
  15. Fyrberg E. A., Goldstein L. S. The Drosophila cytoskeleton. Annu Rev Cell Biol. 1990;6:559–596. doi: 10.1146/annurev.cb.06.110190.003015. [DOI] [PubMed] [Google Scholar]
  16. Hartwig J. H., Chambers K. A., Stossel T. P. Association of gelsolin with actin filaments and cell membranes of macrophages and platelets. J Cell Biol. 1989 Feb;108(2):467–479. doi: 10.1083/jcb.108.2.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Heintzelman M. B., Frankel S. A., Artavanis-Tsakonas S., Mooseker M. S. Cloning of a secretory gelsolin from Drosophila melanogaster. J Mol Biol. 1993 Apr 5;230(3):709–716. doi: 10.1006/jmbi.1993.1191. [DOI] [PubMed] [Google Scholar]
  18. Jackson I. J. A reappraisal of non-consensus mRNA splice sites. Nucleic Acids Res. 1991 Jul 25;19(14):3795–3798. doi: 10.1093/nar/19.14.3795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Janmey P. A., Lamb J., Allen P. G., Matsudaira P. T. Phosphoinositide-binding peptides derived from the sequences of gelsolin and villin. J Biol Chem. 1992 Jun 15;267(17):11818–11823. [PubMed] [Google Scholar]
  20. Karr T. L., Alberts B. M. Organization of the cytoskeleton in early Drosophila embryos. J Cell Biol. 1986 Apr;102(4):1494–1509. doi: 10.1083/jcb.102.4.1494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kellogg D. R., Mitchison T. J., Alberts B. M. Behaviour of microtubules and actin filaments in living Drosophila embryos. Development. 1988 Aug;103(4):675–686. doi: 10.1242/dev.103.4.675. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Kiehart D. P. The actin membrane skeleton in Drosophila development. Semin Cell Biol. 1990 Oct;1(5):325–339. [PubMed] [Google Scholar]
  24. Kwiatkowski D. J., Mehl R., Yin H. L. Genomic organization and biosynthesis of secreted and cytoplasmic forms of gelsolin. J Cell Biol. 1988 Feb;106(2):375–384. doi: 10.1083/jcb.106.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kwiatkowski D. J., Stossel T. P., Orkin S. H., Mole J. E., Colten H. R., Yin H. L. Plasma and cytoplasmic gelsolins are encoded by a single gene and contain a duplicated actin-binding domain. Nature. 1986 Oct 2;323(6087):455–458. doi: 10.1038/323455a0. [DOI] [PubMed] [Google Scholar]
  26. Lassing I., Lindberg U. Evidence that the phosphatidylinositol cycle is linked to cell motility. Exp Cell Res. 1988 Jan;174(1):1–15. doi: 10.1016/0014-4827(88)90136-x. [DOI] [PubMed] [Google Scholar]
  27. Lee W. M., Galbraith R. M. The extracellular actin-scavenger system and actin toxicity. N Engl J Med. 1992 May 14;326(20):1335–1341. doi: 10.1056/NEJM199205143262006. [DOI] [PubMed] [Google Scholar]
  28. McLaughlin P. J., Gooch J. T., Mannherz H. G., Weeds A. G. Structure of gelsolin segment 1-actin complex and the mechanism of filament severing. Nature. 1993 Aug 19;364(6439):685–692. doi: 10.1038/364685a0. [DOI] [PubMed] [Google Scholar]
  29. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Miller K. G., Alberts B. M. F-actin affinity chromatography: technique for isolating previously unidentified actin-binding proteins. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4808–4812. doi: 10.1073/pnas.86.13.4808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Miller K. G., Field C. M., Alberts B. M. Actin-binding proteins from Drosophila embryos: a complex network of interacting proteins detected by F-actin affinity chromatography. J Cell Biol. 1989 Dec;109(6 Pt 1):2963–2975. doi: 10.1083/jcb.109.6.2963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Mount S. M., Burks C., Hertz G., Stormo G. D., White O., Fields C. Splicing signals in Drosophila: intron size, information content, and consensus sequences. Nucleic Acids Res. 1992 Aug 25;20(16):4255–4262. doi: 10.1093/nar/20.16.4255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nodes B. R., Shackelford J. E., Lebherz H. G. Synthesis and secretion of serum gelsolin by smooth muscle tissue. J Biol Chem. 1987 Apr 15;262(11):5422–5427. [PubMed] [Google Scholar]
  34. Pollard T. D., Cooper J. A. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem. 1986;55:987–1035. doi: 10.1146/annurev.bi.55.070186.005011. [DOI] [PubMed] [Google Scholar]
  35. Prendergast G. C., Ziff E. B. Mbh 1: a novel gelsolin/severin-related protein which binds actin in vitro and exhibits nuclear localization in vivo. EMBO J. 1991 Apr;10(4):757–766. doi: 10.1002/j.1460-2075.1991.tb08007.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Reuter R., Scott M. P. Expression and function of the homoeotic genes Antennapedia and Sex combs reduced in the embryonic midgut of Drosophila. Development. 1990 Jun;109(2):289–303. doi: 10.1242/dev.109.2.289. [DOI] [PubMed] [Google Scholar]
  37. Sakurai T., Kurokawa H., Nonomura Y. The Ca2(+)-dependent actin filament-severing activity of 74-kDa protein (adseverin) resides in its NH2-terminal half. J Biol Chem. 1991 Mar 5;266(7):4581–4585. [PubMed] [Google Scholar]
  38. Smith D. B., Janmey P. A., Herbert T. J., Lind S. E. Quantitative measurement of plasma gelsolin and its incorporation into fibrin clots. J Lab Clin Med. 1987 Aug;110(2):189–195. [PubMed] [Google Scholar]
  39. Sprenger F., Nüsslein-Volhard C. Torso receptor activity is regulated by a diffusible ligand produced at the extracellular terminal regions of the Drosophila egg. Cell. 1992 Dec 11;71(6):987–1001. doi: 10.1016/0092-8674(92)90394-r. [DOI] [PubMed] [Google Scholar]
  40. Tautz D., Pfeifle C. A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma. 1989 Aug;98(2):81–85. doi: 10.1007/BF00291041. [DOI] [PubMed] [Google Scholar]
  41. Wang L. L., Spudich J. A. A 45,000-mol-wt protein from unfertilized sea urchin eggs severs actin filaments in a calcium-dependent manner and increases the steady-state concentration of nonfilamentous actin. J Cell Biol. 1984 Sep;99(3):844–851. doi: 10.1083/jcb.99.3.844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Warn R. M., Magrath R. F-actin distribution during the cellularization of the Drosophila embryo visualized with FL-phalloidin. Exp Cell Res. 1983 Jan;143(1):103–114. doi: 10.1016/0014-4827(83)90113-1. [DOI] [PubMed] [Google Scholar]
  43. Way M., Gooch J., Pope B., Weeds A. G. Expression of human plasma gelsolin in Escherichia coli and dissection of actin binding sites by segmental deletion mutagenesis. J Cell Biol. 1989 Aug;109(2):593–605. doi: 10.1083/jcb.109.2.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Way M., Pope B., Gooch J., Hawkins M., Weeds A. G. Identification of a region in segment 1 of gelsolin critical for actin binding. EMBO J. 1990 Dec;9(12):4103–4109. doi: 10.1002/j.1460-2075.1990.tb07632.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Way M., Pope B., Weeds A. G. Are the conserved sequences in segment 1 of gelsolin important for binding actin? J Cell Biol. 1992 Mar;116(5):1135–1143. doi: 10.1083/jcb.116.5.1135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Way M., Weeds A. Nucleotide sequence of pig plasma gelsolin. Comparison of protein sequence with human gelsolin and other actin-severing proteins shows strong homologies and evidence for large internal repeats. J Mol Biol. 1988 Oct 20;203(4):1127–1133. doi: 10.1016/0022-2836(88)90132-5. [DOI] [PubMed] [Google Scholar]
  47. Weeds A., Maciver S. F-actin capping proteins. Curr Opin Cell Biol. 1993 Feb;5(1):63–69. doi: 10.1016/s0955-0674(05)80009-2. [DOI] [PubMed] [Google Scholar]
  48. Young P. E., Pesacreta T. C., Kiehart D. P. Dynamic changes in the distribution of cytoplasmic myosin during Drosophila embryogenesis. Development. 1991 Jan;111(1):1–14. doi: 10.1242/dev.111.1.1. [DOI] [PubMed] [Google Scholar]
  49. Yu F. X., Johnston P. A., Südhof T. C., Yin H. L. gCap39, a calcium ion- and polyphosphoinositide-regulated actin capping protein. Science. 1990 Dec 7;250(4986):1413–1415. doi: 10.1126/science.2255912. [DOI] [PubMed] [Google Scholar]
  50. Yu F. X., Sun H. Q., Janmey P. A., Yin H. L. Identification of a polyphosphoinositide-binding sequence in an actin monomer-binding domain of gelsolin. J Biol Chem. 1992 Jul 25;267(21):14616–14621. [PubMed] [Google Scholar]
  51. von Heijne G. The signal peptide. J Membr Biol. 1990 May;115(3):195–201. doi: 10.1007/BF01868635. [DOI] [PubMed] [Google Scholar]

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