Essential functions and actin-binding surfaces of yeast cofilin revealed by systematic mutagenesis

Pekka Lappalainen, Elena V. Fedorov, Alexander A. Fedorov, Steven C. Almo, David G. Drubin

Research output: Contribution to journalArticle

198 Citations (Scopus)

Abstract

Cofilin stimulates actin filament turnover in vivo. The phenotypes of twenty yeast cofilin mutants generated by systematic mutagenesis were determined. Ten grew as well as the wild type and showed no cytoskeleton defects, seven were recessive-lethal and three were conditional-lethal and caused severe actin organization defects. Biochemical characterization of interactions between nine mutant yeast cofilins and yeast actin provided evidence that F-actin binding and depolymerization are essential cofilin functions. Locating the mutated residues on the yeast cofilin molecular structure allowed several important conclusions to be drawn. First, residues required for actin monomer binding are proximal to each other. Secondly, additional residues are required for interactions with actin filaments; these residues might bind an adjacent subunit in the actin filament. Thirdly, despite striking structural similarity, cofilin interacts with actin in a different manner from gelsolin segment-1. Fourthly, a previously unrecognized cofilin function or interaction is suggested by identification of spatially proximal residues important for cofilin function in vivo, but not for actin interactions in vitro. Finally, mutation of the cofilin N-terminus suggests that its sequence is conserved because of its critical role in actin interactions, not because it is sometimes a target for protein kinases.

Original languageEnglish (US)
Pages (from-to)5520-5530
Number of pages11
JournalEMBO Journal
Volume16
Issue number18
DOIs
StatePublished - Sep 15 1997

Fingerprint

Actin Depolymerizing Factors
Mutagenesis
Yeast
Actins
Yeasts
Actin Cytoskeleton
Gelsolin
Conserved Sequence
Defects
Depolymerization
Molecular Structure
Cytoskeleton
Protein Kinases
Molecular structure
Monomers
Phenotype

Keywords

  • Actin
  • Cofilin
  • Mutagenesis
  • Yeast

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Essential functions and actin-binding surfaces of yeast cofilin revealed by systematic mutagenesis. / Lappalainen, Pekka; Fedorov, Elena V.; Fedorov, Alexander A.; Almo, Steven C.; Drubin, David G.

In: EMBO Journal, Vol. 16, No. 18, 15.09.1997, p. 5520-5530.

Research output: Contribution to journalArticle

Lappalainen, Pekka ; Fedorov, Elena V. ; Fedorov, Alexander A. ; Almo, Steven C. ; Drubin, David G. / Essential functions and actin-binding surfaces of yeast cofilin revealed by systematic mutagenesis. In: EMBO Journal. 1997 ; Vol. 16, No. 18. pp. 5520-5530.
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