Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Feb 1;132(3):335–344. doi: 10.1083/jcb.132.3.335

Overexpression of cofilin stimulates bundling of actin filaments, membrane ruffling, and cell movement in Dictyostelium

PMCID: PMC2120717  PMID: 8636212

Abstract

Cofilin is a low molecular weight actin-modulating protein whose structure and function are conserved among eucaryotes. Cofilin exhibits in vitro both a monomeric actin-sequestering activity and a filamentous actin-severing activity. To investigate in vivo functions of cofilin, cofilin was overexpressed in Dictyostelium discoideum cells. An increase in the content of D. discoideum cofilin (d-cofilin) by sevenfold induced a co-overproduction of actin by threefold. In cells over-expressing d-cofilin, the amount of filamentous actin but not that of monomeric actin was increased. Overexpressed d-cofilin co-sedimented with actin filaments, suggesting that the sequestering activity of d- cofilin is weak in vivo. The overexpression of d-cofilin increased actin bundles just beneath ruffling membranes where d-cofilin was co- localized. The overexpression of d-cofilin also stimulated cell movement as well as membrane ruffling. We have demonstrated in vitro that d-cofilin transformed latticework of actin filaments cross-linked by alpha-actinin into bundles probably by severing the filaments. D. discoideum cofilin may sever actin filaments in vivo and induce bundling of the filaments in the presence of cross-linking proteins so as to generate contractile systems involved in membrane ruffling and cell movement.

Full Text

The Full Text of this article is available as a PDF (2.9 MB).

Selected References

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

  1. Agnew B. J., Minamide L. S., Bamburg J. R. Reactivation of phosphorylated actin depolymerizing factor and identification of the regulatory site. J Biol Chem. 1995 Jul 21;270(29):17582–17587. doi: 10.1074/jbc.270.29.17582. [DOI] [PubMed] [Google Scholar]
  2. Aizawa H., Sutoh K., Tsubuki S., Kawashima S., Ishii A., Yahara I. Identification, characterization, and intracellular distribution of cofilin in Dictyostelium discoideum. J Biol Chem. 1995 May 5;270(18):10923–10932. doi: 10.1074/jbc.270.18.10923. [DOI] [PubMed] [Google Scholar]
  3. Bamburg J. R., Bray D. Distribution and cellular localization of actin depolymerizing factor. J Cell Biol. 1987 Dec;105(6 Pt 1):2817–2825. doi: 10.1083/jcb.105.6.2817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cooper J. A., Blum J. D., Williams R. C., Jr, Pollard T. D. Purification and characterization of actophorin, a new 15,000-dalton actin-binding protein from Acanthamoeba castellanii. J Biol Chem. 1986 Jan 5;261(1):477–485. [PubMed] [Google Scholar]
  5. Davidson M. M., Haslam R. J. Dephosphorylation of cofilin in stimulated platelets: roles for a GTP-binding protein and Ca2+. Biochem J. 1994 Jul 1;301(Pt 1):41–47. doi: 10.1042/bj3010041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Firtel R. A., Silan C., Ward T. E., Howard P., Metz B. A., Nellen W., Jacobson A. Extrachromosomal replication of shuttle vectors in Dictyostelium discoideum. Mol Cell Biol. 1985 Nov;5(11):3241–3250. doi: 10.1128/mcb.5.11.3241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hawkins M., Pope B., Maciver S. K., Weeds A. G. Human actin depolymerizing factor mediates a pH-sensitive destruction of actin filaments. Biochemistry. 1993 Sep 28;32(38):9985–9993. doi: 10.1021/bi00089a014. [DOI] [PubMed] [Google Scholar]
  8. Hayden S. M., Miller P. S., Brauweiler A., Bamburg J. R. Analysis of the interactions of actin depolymerizing factor with G- and F-actin. Biochemistry. 1993 Sep 28;32(38):9994–10004. doi: 10.1021/bi00089a015. [DOI] [PubMed] [Google Scholar]
  9. Iida K., Moriyama K., Matsumoto S., Kawasaki H., Nishida E., Yahara I. Isolation of a yeast essential gene, COF1, that encodes a homologue of mammalian cofilin, a low-M(r) actin-binding and depolymerizing protein. Gene. 1993 Feb 14;124(1):115–120. doi: 10.1016/0378-1119(93)90770-4. [DOI] [PubMed] [Google Scholar]
  10. Kanamori T., Hayakawa T., Suzuki M., Titani K. Identification of two 17-kDa rat parotid gland phosphoproteins, subjects for dephosphorylation upon beta-adrenergic stimulation, as destrin- and cofilin-like proteins. J Biol Chem. 1995 Apr 7;270(14):8061–8067. doi: 10.1074/jbc.270.14.8061. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Maciver S. K., Wachsstock D. H., Schwarz W. H., Pollard T. D. The actin filament severing protein actophorin promotes the formation of rigid bundles of actin filaments crosslinked with alpha-actinin. J Cell Biol. 1991 Dec;115(6):1621–1628. doi: 10.1083/jcb.115.6.1621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Maciver S. K., Zot H. G., Pollard T. D. Characterization of actin filament severing by actophorin from Acanthamoeba castellanii. J Cell Biol. 1991 Dec;115(6):1611–1620. doi: 10.1083/jcb.115.6.1611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Moon A. L., Janmey P. A., Louie K. A., Drubin D. G. Cofilin is an essential component of the yeast cortical cytoskeleton. J Cell Biol. 1993 Jan;120(2):421–435. doi: 10.1083/jcb.120.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Moriyama K., Yonezawa N., Sakai H., Yahara I., Nishida E. Mutational analysis of an actin-binding site of cofilin and characterization of chimeric proteins between cofilin and destrin. J Biol Chem. 1992 Apr 15;267(11):7240–7244. [PubMed] [Google Scholar]
  16. Nagaoka R., Kusano K., Abe H., Obinata T. Effects of cofilin on actin filamentous structures in cultured muscle cells. Intracellular regulation of cofilin action. J Cell Sci. 1995 Feb;108(Pt 2):581–593. doi: 10.1242/jcs.108.2.581. [DOI] [PubMed] [Google Scholar]
  17. Nishida E., Maekawa S., Sakai H. Cofilin, a protein in porcine brain that binds to actin filaments and inhibits their interactions with myosin and tropomyosin. Biochemistry. 1984 Oct 23;23(22):5307–5313. doi: 10.1021/bi00317a032. [DOI] [PubMed] [Google Scholar]
  18. Nishida E., Muneyuki E., Maekawa S., Ohta Y., Sakai H. An actin-depolymerizing protein (destrin) from porcine kidney. Its action on F-actin containing or lacking tropomyosin. Biochemistry. 1985 Nov 5;24(23):6624–6630. doi: 10.1021/bi00344a049. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Quirk S., Maciver S. K., Ampe C., Doberstein S. K., Kaiser D. A., VanDamme J., Vandekerckhove J. S., Pollard T. D. Primary structure of and studies on Acanthamoeba actophorin. Biochemistry. 1993 Aug 24;32(33):8525–8533. doi: 10.1021/bi00084a019. [DOI] [PubMed] [Google Scholar]
  21. Saito T., Lamy F., Roger P. P., Lecocq R., Dumont J. E. Characterization and identification as cofilin and destrin of two thyrotropin- and phorbol ester-regulated phosphoproteins in thyroid cells. Exp Cell Res. 1994 May;212(1):49–61. doi: 10.1006/excr.1994.1117. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Stossel T. P., Chaponnier C., Ezzell R. M., Hartwig J. H., Janmey P. A., Kwiatkowski D. J., Lind S. E., Smith D. B., Southwick F. S., Yin H. L. Nonmuscle actin-binding proteins. Annu Rev Cell Biol. 1985;1:353–402. doi: 10.1146/annurev.cb.01.110185.002033. [DOI] [PubMed] [Google Scholar]
  24. Sun H. Q., Kwiatkowska K., Yin H. L. Actin monomer binding proteins. Curr Opin Cell Biol. 1995 Feb;7(1):102–110. doi: 10.1016/0955-0674(95)80051-4. [DOI] [PubMed] [Google Scholar]
  25. Sutoh K. A transformation vector for dictyostelium discoideum with a new selectable marker bsr. Plasmid. 1993 Sep;30(2):150–154. doi: 10.1006/plas.1993.1042. [DOI] [PubMed] [Google Scholar]
  26. Uyeda T. Q., Ruppel K. M., Spudich J. A. Enzymatic activities correlate with chimaeric substitutions at the actin-binding face of myosin. Nature. 1994 Apr 7;368(6471):567–569. doi: 10.1038/368567a0. [DOI] [PubMed] [Google Scholar]
  27. Yonezawa N., Nishida E., Koyasu S., Maekawa S., Ohta Y., Yahara I., Sakai H. Distribution among tissues and intracellular localization of cofilin, a 21kDa actin-binding protein. Cell Struct Funct. 1987 Oct;12(5):443–452. doi: 10.1247/csf.12.443. [DOI] [PubMed] [Google Scholar]

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

RESOURCES