1. Isolation and characterization of a novel actin filament-binding protein from Saccharomyces cerevisiae
- Author
-
Hiroshi Imamura, Ayako Satoh, Hiroyuki Nakanishi, Takeshi Asakura, Yoshimi Takai, Takuya Sasaki, Hideo Nishioka, Kazuma Tanaka, Hiroshi Obaishi, Fumiko Nagano, and Kazuhiko Hotta
- Subjects
Cancer Research ,Saccharomyces cerevisiae Proteins ,Molecular Sequence Data ,Saccharomyces cerevisiae ,macromolecular substances ,Peptide Mapping ,Open Reading Frames ,Cytosol ,Genetics ,Amino Acid Sequence ,Molecular Biology ,Actin ,chemistry.chemical_classification ,Gel electrophoresis ,Base Sequence ,biology ,Binding protein ,Microfilament Proteins ,biology.organism_classification ,Actins ,Recombinant Proteins ,Amino acid ,Open reading frame ,Biochemistry ,chemistry ,Cytoplasm ,Actin filament binding - Abstract
We purified a novel actin filament (F-actin)-binding protein from the soluble fraction of Saccharomyces cerevisiae by successive column chromatographies by use of the 125I-labeled F-actin blot overlay method. The purified protein showed a minimum Mr of about 140 kDa on SDS-polyacrylamide gel electrophoresis and we named it ABP140. A search with the partial amino acid sequences of ABP140 against the Saccharomyces Genome Database revealed that the open reading frame of the ABP140 gene (ABP140) corresponded to YOR239W fused with YOR240W by the +1 translational frame shift. The encoded protein consisted of 628 amino acids with a calculated Mr of 71,484. The recombinant protein interacted with F-actin and showed the activity to crosslink F-actin into a bundle. Indirect immunofluorescence study demonstrated that ABP140 was colocalized with both cortical actin patches and cytoplasmic actin cables in intact cells. However, elimination of ABP140 by gene disruption did not show a deleterious effect on cell growth or affect the organization of F-actin. These results indicate that ABP140 is not required for cell growth but may be involved in the reorganization of F-actin in the budding yeast.
- Published
- 1998
- Full Text
- View/download PDF