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PMID: 11604420 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Interactions with PIP2, ADP-actin monomers, and capping protein regulate the activity and localization of yeast twinfilin.

The Journal of cell biology ·Vol. 155 ·No. 2 ·2001-10-15 ·Pages 251-60

Palmgren S, Ojala PJ, Wear MA, Cooper JA, Lappalainen P

Abstract

Twinfilin is a ubiquitous actin monomer-binding protein that regulates actin filament turnover in yeast and mammalian cells. To elucidate the mechanism by which twinfilin contributes to actin filament dynamics, we carried out an analysis of yeast twinfilin, and we show here that twinfilin is an abundant protein that localizes to cortical actin patches in wild-type yeast cells. Native gel assays demonstrate that twinfilin binds ADP-actin monomers with higher affinity than ATP-actin monomers. A mutant twinfilin that does not interact with actin monomers in vitro no longer localizes to cortical actin patches when expressed in yeast, suggesting that the ability to interact with actin monomers may be essential for the localization of twinfilin. The localization of twinfilin to the cortical actin cytoskeleton is also disrupted in yeast strains where either the CAP1 or CAP2 gene, encoding for the alpha and beta subunits of capping protein, is deleted. Purified twinfilin and capping protein form a complex on native gels. Twinfilin also interacts with phosphatidylinositol 4,5-bisphosphate (PI[4,5]P2), and its actin monomer-sequestering activity is inhibited by PI(4,5)P2. Based on these results, we propose a model for the biological role of twinfilin as a protein that localizes actin monomers to the sites of rapid filament assembly in cells.

MeSH Terms
Actin Cytoskeleton/metabolism Actin Depolymerizing Factors Actins/metabolism,physiology Adenosine Diphosphate/analogs & derivatives,metabolism,physiology Binding Sites Destrin Microfilament Proteins/genetics,metabolism,physiology Models, Biological Mutation Phosphatidylinositol 4,5-Diphosphate/metabolism,physiology Saccharomyces cerevisiae Proteins Yeasts/metabolism
Chemicals
ADP-G-actin Actin Depolymerizing Factors Actins Destrin Microfilament Proteins Phosphatidylinositol 4,5-Diphosphate Saccharomyces cerevisiae Proteins TWF1 protein, S cerevisiae Adenosine Diphosphate
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Palmgren S
Program in Cellular Biotechnology, Institute of Biotechnology, FIN-00014 University of Helsinki, Helsinki, Finland.
Ojala P J
Wear M A
Cooper J A
Lappalainen P
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200106157
Published
2001-10-15
Epub
2001-00-15
Pages
251-60
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2198831
Subset
IM
Grants
NIGMS NIH HHS · R01 GM047337 · United States
NIGMS NIH HHS · R01 GM047337-09 · United States
External Links
PubMed source DOI: 10.1083/jcb.200106157 PMC Full Text View journal details
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