title: Arabidopsis AtADF1 is functionally affected by mutations on actin binding sites.
abstract: The plant actin depolymerizing factor (ADF) binds to both monomeric and filamentous actin, and is directly involved in the depolymerization of actin filaments. To better understand the actin binding sites of the Arabidopsis thaliana L. AtADF1, we generated mutants of AtADF1 and investigated their functions in vitro and in vivo. Analysis of mutants harboring amino acid substitutions revealed that charged residues (Arg98 and Lys100) located at the α-helix 3 and forming an actin binding site together with the N-terminus are essential for both G- and F-actin binding. The basic residues on the β-strand 5 (K82/A) and the α-helix 4 (R135/A, R137/A) form another actin binding site that is important for F-actin binding. Using transient expression of CFP-tagged AtADF1 mutant proteins in onion (Allium cepa) peel epidermal cells and transgenic Arabidopsis thaliana L. plants overexpressing these mutants, we analyzed how these mutant proteins regulate actin organization and affect seedling growth. Our results show that the ADF mutants with a lower affinity for actin filament binding can still be functional, unless the affinity for actin monomers is also affected. The G-actin binding activity of the ADF plays an essential role in actin binding, depolymerization of actin polymers, and therefore in the control of actin organization.
authors.affiliation: College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, China. chunhai79@yahoo.com
authors.forename: Chun-Hai
authors.initials: CH
authors.lastname: Dong
authors.forename: Wei-Ping
authors.initials: WP
authors.lastname: Tang
authors.forename: Jia-Yao
authors.initials: JY
authors.lastname: Liu
mesh_terms: D051339:Actin Depolymerizing Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D009154:Mutation
chemical_list: D051339:Actin Depolymerizing Factors; D000199:Actins; D029681:Arabidopsis Proteins