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Plant and Cell Physiology, 2003, Vol. 44, No. 10 1088-1099
© 2003 Oxford University Press

Plant 115-kDa Actin-Filament Bundling Protein, P-115-ABP, is a Homologue of Plant Villin and is Widely Distributed in Cells

Etsuo Yokota1,3, Luis Vidali2,4, Motoki Tominaga1,5, Hiroshi Tahara1, Hidefumi Orii1, Yosuke Morizane1, Peter K. Hepler2 and Teruo Shimmen1

1 Department of Life Science, Graduate School of Science, Himeji Institute of Technology, Harima Science Park City, Hyogo, 678-1297 Japan
2 Molecular and Cellular Biology Program, Biology Department, University of Massachusetts, Amherst, MA, U.S.A.

In many cases, actin filaments are arranged into bundles and serve as tracks for cytoplasmic streaming in plant cells. We have isolated an actin-filament bundling protein, which is composed of 115-kDa polypeptide (P-115-ABP), from the germinating pollen of lily, Lilium longiflorum [Nakayasu et al. (1998) Biochem. Biophys. Res. Commun. 249: 61]. P-115-ABP shared similar antigenicity with a plant 135-kDa actin-filament bundling protein (P-135-ABP), a plant homologue of villin. A full-length cDNA clone (ABP115; accession no. AB097407) was isolated from an expression cDNA library of lily pollen by immuno-screening using antisera against P-115-ABP and P-135-ABP. The amino acid sequence of P-115-ABP deduced from this clone showed high homology with those of P-135-ABP and four villin isoforms of Arabidopsis thaliana (AtVLN1, AtVLN2, AtVLN3 and AtVLN4), especially AtVLN4, indicating that P-115-ABP can also be classified as a plant villin. The P-115-ABP isolated biochemically from the germinating lily pollen was able to arrange F-actin filaments with uniform polarity into bundles and this bundling activity was suppressed by Ca2+-calmodulin (CaM), similar to the actin-filament bundling properties of P-135-ABP. The P-115-ABP type of plant villin was widely distributed in plant cells, from algae to land plants. In root hair cells of Hydrocharis dubia, this type of plant villin was co-localized with actin-filament bundles in the transvacuolar strands and the sub-cortical regions. Microinjection of the antiserum against P-115-ABP into living root hair cells caused the disappearance of transvaculor strands and alteration of the route of cytoplasmic streaming. In internodal cells of Chara corallina in which the P-135-ABP type of plant villin is lacking, the P-115-ABP type showed co-localization with actin-filament cables anchored on the intracellular surface of chloroplasts. These results indicated that plant villins are widely distributed and involved in the organization of actin filaments into bundles throughout the plant kingdom.

3 Corresponding author: E-mail, yokota{at}sci.himeji-tech.ac.jp; Fax, +81-791-58-0175.

4 Present address: Brigham and Women’s Hospital Division of Hematology, LMR Center 301, 221 Longwood Avenue, Boston, MA 02115, U.S.A.

5 Present address: Kansai Advanced Research Center, Communications Research Laboratory, Kobe, 651-2492 Japan


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