Structural plasticity of functional actin: pictures of actin binding protein and polymer interfaces.
Actin is one of the most conserved and versatile proteins capable of forming homopolymers and interacting with numerous other proteins in the cell. We performed an alanine mutagenesis scan covering the entire beta-actin molecule. Somewhat surprisingly, the majority of the mutants were capable of reaching a stable conformation. We tested the ability of these mutants to bind to various actin binding proteins, thereby mapping different interfaces with actin. Additionally, we tested their ability to copolymerize with alpha-actin in order to localize regions in actin that contact neighboring protomers in the filament. Hereby, we could discriminate between two existing models for filamentous actin and our data strongly support the right-handed double-stranded helix model. We present data corroborating this model in vivo. Mutants defective in copolymerization do not colocalize with the actin cytoskeleton and some impair its normal function, thereby disturbing cell shape.
Rommelaere
Heidi
H
Department of Biochemistry and Molecular Biology, Faculty of Medicine and Health Sciences, Ghent University, A. Baertsoenkaai 3, B-9000, Gent, Belgium. heidi.rommelaere@ugent.be
Waterschoot
Davy
D
Neirynck
Katrien
K
Vandekerckhove
Joël
J
Ampe
Christophe
C
D000199:Actins / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism*; D009154:Mutation; D017434:Protein Structure, Tertiary; D013947:Thymosin / Q000378:metabolism; D000099201:Thymosin beta(4)
D000199:Actins; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D013947:Thymosin; D000099201:Thymosin beta(4)