Isolation and characterization of a novel actin filament-binding protein from Saccharomyces cerevisiae (original) (raw)

Oncogene volume 16, pages 121–130 (1998)Cite this article

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 **M**r 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.

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Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry, Osaka University Medical School, Suita, 565, Japan
    Takeshi Asakura, Takuya Sasaki, Fumiko Nagano, Hiroshi Imamura, Kazuhiko Hotta, Kazuma Tanaka & Yoshimi Takai
  2. Takai Biotimer Project, ERATO, Japan Science and Technology Corporation, c/o JCR Pharmaceuticals Co., Ltd., 2-2-10 Murotani, Nishi-ku, 651-22, Kobe, Japan
    Ayako Satoh, Hiroshi Obaishi, Hideo Nishioka, Hiroyuki Nakanishi & Yoshimi Takai

Authors

  1. Takeshi Asakura
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  2. Takuya Sasaki
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  3. Fumiko Nagano
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  4. Ayako Satoh
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  5. Hiroshi Obaishi
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  6. Hideo Nishioka
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  7. Hiroshi Imamura
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  8. Kazuhiko Hotta
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  9. Kazuma Tanaka
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  10. Hiroyuki Nakanishi
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  11. Yoshimi Takai
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Asakura, T., Sasaki, T., Nagano, F. et al. Isolation and characterization of a novel actin filament-binding protein from Saccharomyces cerevisiae.Oncogene 16, 121–130 (1998). https://doi.org/10.1038/sj.onc.1201487

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