Plant Molecular Biology

, Volume 45, Issue 5, pp 517–527 | Cite as

Molecular identification and characterization of the Arabidopsis AtADF1, AtADF5 and AtADF6 genes

  • Chun-Hai Dong
  • Benedikt Kost
  • Guixian Xia
  • Nam-Hai Chua


Actin depolymerizing factor (ADF) is a key regulator of the organization of the actin cytoskeleton during various cellular activities. We found that ADF genes in Arabidopsis form a large family consisting of at least nine members, four of which were cloned and sequenced in this study. Comparison of genomic and cDNA sequences showed that the AtADF1, AtADF5, and AtADF6 genes all contain two introns at conserved positions. Analysis of transgenic Arabidopsis plants carrying promoter-GUS fusion constructs revealed that AtADF1 and AtADF6 are expressed in the vascular tissues of all organs, whereas expression of AtADF5 is restricted to the root tip meristem. GFP-AtADF1, GFP-AtADF5, and GFP-AtADF6 fusion proteins were found to bind to actin filaments in vivo, and to reorganize the actin cytoskeleton when transiently expressed in plant cells.

ADF (actin depolymerizing factor) actin cytoskeleton Arabidopsis thaliana GUS GFP 


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  1. Agnew, B.J., Minamide, L.S. and Bamburg, JR. 1995. Reactivation of phosphorylated actin depolymerizing factor and identification of the regulatory site. J. Biol. Chem. 270: 17582–17587.Google Scholar
  2. Aizawa, H., Sutoh, K., Tsubuki, S., Kawashima, S., Ishii, A. and Yahara, I. 1995. Identification, characterization, and intracellu-lar distribution of cofilin in Dictyostelium discoideum. J. Biol. Chem. 270: 10923–10932.Google Scholar
  3. Aizawa, H., Sutoh, K. and Yahara, I. 1996. Overexpression of cofilin stimulates bundling of actin filaments, membrane ruffling, and cell movement in Dictyostelium. J. Cell Biol. 132: 335–344.Google Scholar
  4. Aizawa, H., Fukui, Y. and Yahara, I. 1997. Live dynamics of Dic-tyostelium cofilin suggests a role in remodeling actin latticework into bundles. J. Cell Sci. 110: 2333–2344.Google Scholar
  5. Ayscough, K.R 1998. In vivo functions of actin-binding proteins. Curr. Opin. Cell Biol. 10: 102–111.Google Scholar
  6. Barnes, S.A., Nishizawa, N.K., Quaggi, R.B., Whitelam, G.C. and Chua, N.-H. 1996. Far-red light blocks greening of Arabidopsis seedling via a phytochrome A-mediated change in plastid change in plastic development. Plant Cell 8: 601–615.Google Scholar
  7. Christensen, H.E.M., Ramachandran, S., Tan, C.T., Surana, U., Dong, C.-H. and Chua, N.-H. 1996. Arabidopsis profilins are functionally similar to yeast profilins: identification of vascular bundle-specific profilin and a pollen-specific profilin.Plant J. 10: 269–279.Google Scholar
  8. Carlier, M.F., Santolini, J., Lanrent, V., Melki, R., Didry, D., Hong, Y., Xia, G.-X., Chua N.-H. and Pantolni, D. 1997. Actin de-polymerizing factor (ADF/cofilin) enhances the rate of filament turnover: implication in actin-based motility. J. Cell Biol. 136: 1307–1322.Google Scholar
  9. Carlier, M.F. 1998. Control of actin dynamics. Curr. Opin. Cell Biol. 10: 45–51.Google Scholar
  10. Eun, S.-O. and Lee Y. 1997. Actin filaments of guard cells are reor-ganized in response to light and abscisic acid. Plant Physiol. 115: 1491–1498.Google Scholar
  11. Fedorov, A.A., Lappalainen, P., Fedorov, E.V., Drubin, D.G. and Almo, S.C. 1997. Structure determination of yeast cofilin. Nature Struct. Biol. 4: 366–369.Google Scholar
  12. Goode, B.L., Drubin, D.G. and Lappalainen, P. 1998. Regulation of the cortical actin cytoskeleton in budding yeast by Twinfilin, a ubiquitous actin monomer-sequestering protein. J. Cell Biol. 142: 723–733.Google Scholar
  13. Gunsalus, K.C., Bonaccorsi, S., Williams, E., Verni, F., Gatti, M. and Goldberg, M.L. 1995. Mutations in twinstar,aDrosophila gene encoding a cofilin/ADF homologue, result in defects in centrosome migration and cytokinesis. J. Cell Biol. 131: 1243–1259..527Google Scholar
  14. Hatanaka, H., Ogura, K., Moriyama, M., Ichikawa, S., Yahara, I. and Inagaki, F. 1996. Tertiary structure of destrin and structural similarity between two actin-regulating protein families. Cell 85: 1047–1055.Google Scholar
  15. Hussey, P.J., Yuan, M., Calder, G., Khan, S. and Lioyd, C.W. 1998. Microinjection of pollen-specific actin-depolymerizing factor, ZmADF1, reorients F-actin strands in Tradescantia stamen hair cells. Plant J. 14: 353–357.Google Scholar
  16. Huang, S., McDowell, J.M., Weise, M.J. and Meagher, R.B. 1996. The Arabidopsis profilin gene family: evidence for an ancient split between constitutive and pollen-specific profilin genes. Plant Physiol. 111: 115–126.Google Scholar
  17. Iida, K., Moriyama, K., Matsumoto, S., Kawasaki, H., Nishida, E. and Yahara, I. 1993. Isolation of a yeast essential gene, COF1, that encodes a homologue of mammalian cofilin, a low-Mr actin-binding and depolymerizing protein. Gene 124: 115–120.Google Scholar
  18. Jefferson, R.A. 1987. Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5: 387–405.Google Scholar
  19. Johnston, G., Prendergast, J. and Singer, R. 1991. The Saccha-romyces cerevisiae MYO2 gene encodes an essential myosin for vectorial transport of vesicles. I. Cell Biol. 113: 539–551.Google Scholar
  20. Kim, S.-R., Kim, Y. and An, G. 1993. Molecular cloning and characterization of anther-preferential cDNA encoding a putative actin-depolymerizing factor. Plant Mol. Biol. 21: 39–45.Google Scholar
  21. Klahre, U., Friederich, E., Kost, B., Louvard, D. and Chua, N.-H. 1999. The actin binding protein VILLIN is expressed ubiquitously in Arabidopsis. Plant Physiol. 122: 35–47.Google Scholar
  22. Kopczak, S.D., Haas, N.A., Hussey, P.J., Silflow, C.D. and Snustad, D.P. 1992. The small genome of Arabidopsis contains at least six expressed alpha-tubulin genes. Plant Cell 4: 539–547.Google Scholar
  23. Kost, B., Spielhofer, P. and Chua, N.-H. 1998. A GFP-mouse talin fusion protein labels plant actin filaments in vivo and visualizes the actin cytoskeleton in growing pollen tubes. Plant J. 16: 393–401.Google Scholar
  24. Kost, B., Mathur, J. and Chua, N.-H. 1999. Cytoskeleton in plant development. Curr. Opin. Plant Biol. 2: 462–470.Google Scholar
  25. Kost, B., Spielholfer, P., Mathur, J., Dong, C.-H. and Chua, N.-H. 2000. Non-invasive F-actin visualization in living plant cells using a GFP-mouse talin fusion protein. In: C.J. Staiger, F. Baluska, D. Volkmann and P. Barlow (Eds.) Actin: A Dynamic Framework For Multiple Plant Cell Functions, Kluwer Academic Publishers, Dordrecht, Netherlands, pp. 637–659.Google Scholar
  26. Lappalainen, P. and Drubin, D.G. 1997. Cofilin promotes rapid actin filament turnover in vivo. Nature 388: 78–82.Google Scholar
  27. Leonard, S., Gittis, A., Petrella, E., Pollard, T. and Lattman, E. 1997. Crystal structure of the actin-binding protein actophorin from Acanthamoeba. Nature Struct. Biol. 4: 369–373.Google Scholar
  28. Lopez, I., Anthony, R.G., Maciver, S.K., Jiang, C.-J., Khan, S,, Weeds, A.G. and Hussey, P.J.1996. Pollen specific expression of maize genes encoding actin depolymerizing factor-like proteins. Proc. Natl. Acad. Sci. USA 93: 7415–7420.Google Scholar
  29. Mabuchi, 1.1983. An actin-depolymerizing protein (depactin) from starfish oocytes: properties and interaction with actin. J. Cell Biol. 97: 1612–1621.Google Scholar
  30. Maciver, S.K., Pope, B.J., Whytock, S. and Weeds, A.G. 1998. The effect of two actin depolymerizing factors (ADF/cofilins) on actin filament turnover: pH sensitivity of F-actin binding by hu-man ADF, but not of Acanthamoeba actophorin. Eur. J. Biochem. 256: 388–397.Google Scholar
  31. Marthur, J., Spielhofer, P., Kost, B. and Chua, N.-H. 1999. The actin cytoskeleton is required to elaborate and maintain spatial patterning during trichome cell morphogenesis in Arabidopsis thaliana. Development 126: 5559–5568.Google Scholar
  32. Meagher, R.B., McKinney, E.C. and Kandasmy, M.K. 1999. Iso-variant dynamics expand and buffer the responses of complex systems: the diverse plant actin gene family. Plant Cell 11: 995–1005.Google Scholar
  33. Mckinney, E.C. and Meagher, R.B. 1998. Members of the Ara-bidopsis actin gene family are widely dispersed in the genome. Genetics 149: 663–675.Google Scholar
  34. McGough, A., Pope, B., Chiu, W. and Weeds, A. 1997. Cofilin changes the twist of F-actin: implications for actin filament dynamics and cellular function. J. Cell Biol. 13: 771–781.Google Scholar
  35. Miller, D.D., Ruijter, N.C.A., Bisseling, T. and Emons, A.M.C. 1998. The role of actin in root hair morphogenesis: studies with lipochito-oligosaccharide as a growth stimulator and cytocha-lasin as an actin perturbing drug. Plant J. 17: 141–154.Google Scholar
  36. Moon, A., Janmey, P.A., Loui, K.A. and Drubin, D.G. 1993. Cofilin is an important component of the yeast cortical cytoskeleton. J. Cell Biol. 120: 421–435.Google Scholar
  37. Moriyama, K. and Yahara, I. 1999. Two activities of cofilin, severing and accelerating directional depolymerization of actin filaments, are affected differentially by mutations around the actin-binding helix. EMBO J. 18: 6752–6761.Google Scholar
  38. Moriyama, K., Iida, K. and Yahara, I. 1996. Phosphorylation of Ser-3 of cofilin regulates its essential function on actin. Genes Cells 1: 73–86.Google Scholar
  39. Nagaoka, R., Abe, H., Kusano, K. and Obinata, T. 1995a. Concentration of cofilin, a small actin-binding protein, at the cleavage furrow during cytokinesis. Cell Motil. Cytoskel. 30: 1–7.Google Scholar
  40. Nagaoka, R., Kusano, K.I., Abe, H. and Obinata, T. 1995b. Effects of cofilin on actin filamentous structures in cultured muscle cells. J. Cell Sci. 108: 581–593.Google Scholar
  41. Rosenblatt, J., Agnew, B.J., Abe, H., Bamburg, J.R. and Mitchison, T.J. 1997. Xenopus actin depolymerizing factor/cofilin (XAC) is responsible for the turnover of actin filaments in Listeria monocytogenes tails. J. Cell Biol. 136: 1323–1332.Google Scholar
  42. Rozycka, M., Khan, S., Lopez, I., Greenland, A.J. and Hussey, P.J. 1995. A Zea mays pollen cDNA encoding a putative actin-depolymerizing factor. Plant Physiol. 107: 1011–1012.Google Scholar
  43. Sambrook, D.N., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd. ed. Cold Spring Harbor Laboratory Press, Plainview, NY.Google Scholar
  44. Smertenko, A.P., Jiang, C.-J., Simmons, N.J., Weeds, A.G., Davies, D.R. and Hussey, P.J. 1998. Ser6 in the maize actin-depolymerizing factor, ZmADF3, is phosphorylated by calcium-stimulated protein kinase and is essential for the control of functional activity. Plant J. 14: 187–193.Google Scholar
  45. Staiger, C.J., Gibbon, B.C., Kovar, D.R. and Zonia, L.E. 1997. Profilin and actin-depolymerizing factor: modulators of actin organization in plants. Trends Plant Sci. 2(7): 275–281.Google Scholar
  46. Snustad, D.P., Haas, N.A., Kopczak, S.D. and Silflow, C.D. 1992. The small genome of Arabidopsis contains at least nine expressed beta-tubulin genes. Plant Cell 4: 549–556.Google Scholar
  47. Szymanski, D.B., Marks, M.D., and Wick, S.M. 1999. Organized F-actin is essential for normal trichome morphogenesis in Arabidopsis. Plant Cell 11: 2331–2347.Google Scholar
  48. Valvekens, D., Van Montagu, M. and Lijsebettens, M. 1988. Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants using kanamycin selection. Proc. Natl. Acad. Sci. USA 85: 5536–5540.Google Scholar
  49. Wang, H., Lockwood, S.K., Hoeltzel, M.F. and Schiefelbein. J.W. 1997. The ROOT HAIR DEFECTIVE3 gene encodes an evolutionarily conserved protein with GTP-binding motifs and is required for regulated cell enlargement in Arabidopsis. Genes Dev. 11: 799–811.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Chun-Hai Dong
    • 1
  • Benedikt Kost
    • 1
  • Guixian Xia
    • 1
  • Nam-Hai Chua
    • 2
  1. 1.Laboratory of Plant Cell Biology, Institute of Molecular AgrobiologyNational University of SingaporeSingapore 
  2. 2.Laboratory of Plant Molecular BiologyRockefeller UniversityNew YorkUSA

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