Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Dec 2;127(6):1995–2008. doi: 10.1083/jcb.127.6.1995

Functional differences between L- and T-plastin isoforms

PMCID: PMC2120298  PMID: 7806577

Abstract

Fimbrins/plastins are a family of highly conserved actin-bundling proteins. They are present in all eukaryotic cells including yeast, but each isoform displays a remarkable tissue specificity. T-plastin is normally found in epithelial and mesenchymal cells while L-plastin is present in hematopoietic cells. However, L-plastin has been also found in tumor cells of non-hematopoietic origin (Lin, C.-S., R. H. Aebersold, S. B. Kent, M. Varma, and J. Leavitt. 1988. Mol. Cell. Biol. 8:4659-4668; Lin, C.-S., R. H. Aebersold, and J. Leavitt. 1990. Mol. Cell. Biol. 10: 1818-1821). To learn more about the biological significance of their tissue specificity, we have overproduced the T- and L-plastin isoforms in a fibroblast-like cell line, CV-1, and in a polarized epithelial cell line, LLC-PK1. In CV-1 cells, overproduction of T- and L-plastins induces cell rounding and a concomitant reorganization of actin stress fibers into geodesic structures. L- plastin remains associated with microfilaments while T-plastin is almost completely extracted after treatment of the cells with non-ionic detergent. In LLC-PK1 cells, T-plastin induces shape changes in microvilli and remains associated with microvillar actin filaments after detergent extraction while L-plastin has no effect on these structures and is completely extracted. The effect of T-plastin on the organization of microvilli differs from that of villin, another actin- bundling protein. Our experiments indicate that these two isoforms play differing roles in actin filament organization, and do so in a cell type-specific fashion. Thus it is likely that these plastin isoforms play fundamentally different roles in cell function.

Full Text

The Full Text of this article is available as a PDF (5.6 MB).

Selected References

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

  1. Adams A. E., Botstein D., Drubin D. G. A yeast actin-binding protein is encoded by SAC6, a gene found by suppression of an actin mutation. Science. 1989 Jan 13;243(4888):231–233. doi: 10.1126/science.2643162. [DOI] [PubMed] [Google Scholar]
  2. Adams A. E., Botstein D., Drubin D. G. Requirement of yeast fimbrin for actin organization and morphogenesis in vivo. Nature. 1991 Dec 5;354(6352):404–408. doi: 10.1038/354404a0. [DOI] [PubMed] [Google Scholar]
  3. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bretscher A. Fimbrin is a cytoskeletal protein that crosslinks F-actin in vitro. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6849–6853. doi: 10.1073/pnas.78.11.6849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bretscher A. Fimbrin is a cytoskeletal protein that crosslinks F-actin in vitro. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6849–6853. doi: 10.1073/pnas.78.11.6849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bretscher A., Weber K. Fimbrin, a new microfilament-associated protein present in microvilli and other cell surface structures. J Cell Biol. 1980 Jul;86(1):335–340. doi: 10.1083/jcb.86.1.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bretscher A., Weber K. Villin is a major protein of the microvillus cytoskeleton which binds both G and F actin in a calcium-dependent manner. Cell. 1980 Jul;20(3):839–847. doi: 10.1016/0092-8674(80)90330-x. [DOI] [PubMed] [Google Scholar]
  8. Bretscher A., Weber K. Villin: the major microfilament-associated protein of the intestinal microvillus. Proc Natl Acad Sci U S A. 1979 May;76(5):2321–2325. doi: 10.1073/pnas.76.5.2321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  10. Carley W. W., Bretscher A., Webb W. W. F-actin aggregates in transformed cells contain alpha-actinin and fimbrin but apparently lack tropomyosin. Eur J Cell Biol. 1986 Jan;39(2):313–320. [PubMed] [Google Scholar]
  11. Chambers C., Grey R. D. Development of the structural components of the brush border in absorptive cells of the chick intestine. Cell Tissue Res. 1979;204(3):387–405. doi: 10.1007/BF00233651. [DOI] [PubMed] [Google Scholar]
  12. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  14. Coudrier E., Kerjaschki D., Louvard D. Cytoskeleton organization and submembranous interactions in intestinal and renal brush borders. Kidney Int. 1988 Sep;34(3):309–320. doi: 10.1038/ki.1988.183. [DOI] [PubMed] [Google Scholar]
  15. Coudrier E., Reggio H., Louvard D. Characterization of an integral membrane glycoprotein associated with the microfilaments of pig intestinal microvilli. EMBO J. 1983;2(3):469–475. doi: 10.1002/j.1460-2075.1983.tb01446.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Einberger H., Mertz R., Hofschneider P. H., Neubert W. J. Purification, renaturation, and reconstituted protein kinase activity of the Sendai virus large (L) protein: L protein phosphorylates the NP and P proteins in vitro. J Virol. 1990 Sep;64(9):4274–4280. doi: 10.1128/jvi.64.9.4274-4280.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Friederich E., Huet C., Arpin M., Louvard D. Villin induces microvilli growth and actin redistribution in transfected fibroblasts. Cell. 1989 Nov 3;59(3):461–475. doi: 10.1016/0092-8674(89)90030-5. [DOI] [PubMed] [Google Scholar]
  19. Friederich E., Pringault E., Arpin M., Louvard D. From the structure to the function of villin, an actin-binding protein of the brush border. Bioessays. 1990 Sep;12(9):403–408. doi: 10.1002/bies.950120902. [DOI] [PubMed] [Google Scholar]
  20. Friederich E., Vancompernolle K., Huet C., Goethals M., Finidori J., Vandekerckhove J., Louvard D. An actin-binding site containing a conserved motif of charged amino acid residues is essential for the morphogenic effect of villin. Cell. 1992 Jul 10;70(1):81–92. doi: 10.1016/0092-8674(92)90535-k. [DOI] [PubMed] [Google Scholar]
  21. Geiger B., Tokuyasu K. T., Dutton A. H., Singer S. J. Vinculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4127–4131. doi: 10.1073/pnas.77.7.4127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Glenney J. R., Jr, Kaulfus P., Matsudaira P., Weber K. F-actin binding and bundling properties of fimbrin, a major cytoskeletal protein of microvillus core filaments. J Biol Chem. 1981 Sep 10;256(17):9283–9288. [PubMed] [Google Scholar]
  23. Glück U., Kwiatkowski D. J., Ben-Ze'ev A. Suppression of tumorigenicity in simian virus 40-transformed 3T3 cells transfected with alpha-actinin cDNA. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):383–387. doi: 10.1073/pnas.90.2.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Goldstein D., Djeu J., Latter G., Burbeck S., Leavitt J. Abundant synthesis of the transformation-induced protein of neoplastic human fibroblasts, plastin, in normal lymphocytes. Cancer Res. 1985 Nov;45(11 Pt 2):5643–5647. [PubMed] [Google Scholar]
  25. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  26. Heintzelman M. B., Mooseker M. S. Assembly of the intestinal brush border cytoskeleton. Curr Top Dev Biol. 1992;26:93–122. doi: 10.1016/s0070-2153(08)60442-1. [DOI] [PubMed] [Google Scholar]
  27. Ingber D. E. Cellular tensegrity: defining new rules of biological design that govern the cytoskeleton. J Cell Sci. 1993 Mar;104(Pt 3):613–627. doi: 10.1242/jcs.104.3.613. [DOI] [PubMed] [Google Scholar]
  28. Kreis T. E. Microinjected antibodies against the cytoplasmic domain of vesicular stomatitis virus glycoprotein block its transport to the cell surface. EMBO J. 1986 May;5(5):931–941. doi: 10.1002/j.1460-2075.1986.tb04306.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kreis T. E. Microtubules containing detyrosinated tubulin are less dynamic. EMBO J. 1987 Sep;6(9):2597–2606. doi: 10.1002/j.1460-2075.1987.tb02550.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kübler E., Riezman H. Actin and fimbrin are required for the internalization step of endocytosis in yeast. EMBO J. 1993 Jul;12(7):2855–2862. doi: 10.1002/j.1460-2075.1993.tb05947.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  32. Lazarides E. Actin, alpha-actinin, and tropomyosin interaction in the structural organization of actin filaments in nonmuscle cells. J Cell Biol. 1976 Feb;68(2):202–219. doi: 10.1083/jcb.68.2.202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lin C. S., Aebersold R. H., Kent S. B., Varma M., Leavitt J. Molecular cloning and characterization of plastin, a human leukocyte protein expressed in transformed human fibroblasts. Mol Cell Biol. 1988 Nov;8(11):4659–4668. doi: 10.1128/mcb.8.11.4659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Lin C. S., Aebersold R. H., Leavitt J. Correction of the N-terminal sequences of the human plastin isoforms by using anchored polymerase chain reaction: identification of a potential calcium-binding domain. Mol Cell Biol. 1990 Apr;10(4):1818–1821. doi: 10.1128/mcb.10.4.1818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Lin C. S., Park T., Chen Z. P., Leavitt J. Human plastin genes. Comparative gene structure, chromosome location, and differential expression in normal and neoplastic cells. J Biol Chem. 1993 Feb 5;268(4):2781–2792. [PubMed] [Google Scholar]
  36. Lin C. S., Shen W., Chen Z. P., Tu Y. H., Matsudaira P. Identification of I-plastin, a human fimbrin isoform expressed in intestine and kidney. Mol Cell Biol. 1994 Apr;14(4):2457–2467. doi: 10.1128/mcb.14.4.2457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Matsudaira P. Modular organization of actin crosslinking proteins. Trends Biochem Sci. 1991 Mar;16(3):87–92. doi: 10.1016/0968-0004(91)90039-x. [DOI] [PubMed] [Google Scholar]
  38. Matsushima K., Shiroo M., Kung H. F., Copeland T. D. Purification and characterization of a cytosolic 65-kilodalton phosphoprotein in human leukocytes whose phosphorylation is augmented by stimulation with interleukin 1. Biochemistry. 1988 May 17;27(10):3765–3770. doi: 10.1021/bi00410a037. [DOI] [PubMed] [Google Scholar]
  39. Maunoury R., Robine S., Pringault E., Huet C., Guénet J. L., Gaillard J. A., Louvard D. Villin expression in the visceral endoderm and in the gut anlage during early mouse embryogenesis. EMBO J. 1988 Nov;7(11):3321–3329. doi: 10.1002/j.1460-2075.1988.tb03203.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Messier J. M., Shaw L. M., Chafel M., Matsudaira P., Mercurio A. M. Fimbrin localized to an insoluble cytoskeletal fraction is constitutively phosphorylated on its headpiece domain in adherent macrophages. Cell Motil Cytoskeleton. 1993;25(3):223–233. doi: 10.1002/cm.970250303. [DOI] [PubMed] [Google Scholar]
  41. Meyer R. K., Aebi U. Bundling of actin filaments by alpha-actinin depends on its molecular length. J Cell Biol. 1990 Jun;110(6):2013–2024. doi: 10.1083/jcb.110.6.2013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Namba Y., Ito M., Zu Y., Shigesada K., Maruyama K. Human T cell L-plastin bundles actin filaments in a calcium-dependent manner. J Biochem. 1992 Oct;112(4):503–507. doi: 10.1093/oxfordjournals.jbchem.a123929. [DOI] [PubMed] [Google Scholar]
  43. Pacaud M., Derancourt J. Purification and further characterization of macrophage 70-kDa protein, a calcium-regulated, actin-binding protein identical to L-plastin. Biochemistry. 1993 Apr 6;32(13):3448–3455. doi: 10.1021/bi00064a031. [DOI] [PubMed] [Google Scholar]
  44. Pacaud M., Harricane M. C. Calcium control of macrophage cytoplasmic gelation: evidence for the involvement of the 70,000 Mr actin-bundling protein. J Cell Sci. 1987 Aug;88(Pt 1):81–94. doi: 10.1242/jcs.88.1.81. [DOI] [PubMed] [Google Scholar]
  45. Rodríguez Fernández J. L., Geiger B., Salomon D., Ben-Ze'ev A. Overexpression of vinculin suppresses cell motility in BALB/c 3T3 cells. Cell Motil Cytoskeleton. 1992;22(2):127–134. doi: 10.1002/cm.970220206. [DOI] [PubMed] [Google Scholar]
  46. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  47. Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
  48. Weber A., Nachmias V. T., Pennise C. R., Pring M., Safer D. Interaction of thymosin beta 4 with muscle and platelet actin: implications for actin sequestration in resting platelets. Biochemistry. 1992 Jul 14;31(27):6179–6185. doi: 10.1021/bi00142a002. [DOI] [PubMed] [Google Scholar]
  49. Zu Y. L., Shigesada K., Nishida E., Kubota I., Kohno M., Hanaoka M., Namba Y. 65-kilodalton protein phosphorylated by interleukin 2 stimulation bears two putative actin-binding sites and two calcium-binding sites. Biochemistry. 1990 Sep 11;29(36):8319–8324. doi: 10.1021/bi00488a017. [DOI] [PubMed] [Google Scholar]
  50. de Arruda M. V., Watson S., Lin C. S., Leavitt J., Matsudaira P. Fimbrin is a homologue of the cytoplasmic phosphoprotein plastin and has domains homologous with calmodulin and actin gelation proteins. J Cell Biol. 1990 Sep;111(3):1069–1079. doi: 10.1083/jcb.111.3.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES