Journal Article

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

Etsuo Yokota, Luis Vidali, Motoki Tominaga, Hiroshi Tahara, Hidefumi Orii, Yosuke Morizane, Peter K. Hepler and Teruo Shimmen

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 44, issue 10, pages 1088-1099
Published in print October 2003 | ISSN: 0032-0781
Published online October 2003 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcg132
Plant 115-kDa Actin-Filament Bundling Protein, P-115-ABP, is a Homologue of Plant Villin and is Widely Distributed in Cells

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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.

Keywords: Keywords: Actin — Actin-filament bundling protein — Lily — Pollen tube — Villin.; Abbreviations: ABP, actin-filament bundling protein; BSA, bovine serum albumin; CaM, calmodulin; CBB, Coomassie brilliant blue; P-115-ABP, plant 115-kDa actin-filament bundling protein; P-135-ABP, plant 135-kDa actin-filament bundling protein; PBS, phosphate buffered saline; RP, rohdamine-phalloidin; S-1, myosin subfragment-1; TCA, trichloroacetic acid.

Journal Article.  7117 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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