ISOLATION AND CHARACTERIZATION OF A NOVEL ACTIN FILAMENT-BINDING PROTEIN FROM SACCHAROMYCES-CEREVISIAE

Citation
T. Asakura et al., ISOLATION AND CHARACTERIZATION OF A NOVEL ACTIN FILAMENT-BINDING PROTEIN FROM SACCHAROMYCES-CEREVISIAE, Oncogene, 16(1), 1998, pp. 121-130
Citations number
50
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Oncology,Biology,"Cell Biology","Genetics & Heredity
Journal title
ISSN journal
0950-9232
Volume
16
Issue
1
Year of publication
1998
Pages
121 - 130
Database
ISI
SICI code
0950-9232(1998)16:1<121:IACOAN>2.0.ZU;2-#
Abstract
We purified a novel actin filament (F-actin)-binding protein from the soluble fraction of Saccharomyces cerevisiae by successive column chro matographies by use of the I-125-labeled F-actin blot overlay method. The purified protein showed a minimum M-r of about 140 kDa on SDS-poly acrylamide gel electrophoresis and we named it ABP140. A search with t he partial amino acid sequences of ABP140 against the Saccharomyces Ge nome Database revealed that the open reading frame of the ABP140 gene (ABP140) corresponded to YOR239W fused with YOR240W by the +1 translat ional frame shift. The encoded protein consisted of 628 amino acids wi th 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. I ndirect immunofluorescence study demonstrated that ABP140 was colocali zed with both cortical actin patches and cytoplasmic actin cables in i ntact cells. However, elimination of ABP140 by gene disruption did not show a deleterious effect on cell growth or affect the organization o f 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 bu dding yeast.