Fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin Cdc12

Andrew J. Bestul, Jenna R. Christensen, Agnieszka P. Grzegorzewska, Thomas A. Burke, Jennifer A. Sees, Robert T. Carroll, Vladimir Sirotkin, Robert J. Keenan, David R. Kovar*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

The evolutionarily conserved small actin-monomer binding protein profilin is believed to be a housekeeping factor that maintains a general pool of unassembled actin. However, despite similar primary sequences, structural folds, and affinities for G-Actin and poly-l-proline, budding yeast profilin ScPFY fails to complement fission yeast profilin SpPRF temperature-sensitive mutant cdc3-124 cells. To identify profilin's essential properties, we built a combinatorial library of ScPFY variants containing either WT or SpPRF residues at multiple positions and carried out a genetic selection to isolate variants that support life in fission yeast. We subsequently engineered ScPFY(9-Mut), a variant containing nine substitutions in the actin-binding region, which complements cdc3-124 cells. ScPFY(9-Mut), but not WT ScPFY, suppresses severe cytokinesis defects in cdc3-124 cells. Furthermore, the major activity rescued by ScPFY(9-Mut) is the ability to enhance cytokinesis formin Cdc12-mediated actin assembly in vitro, which allows cells to assemble functional contractile rings. Therefore an essential role of profilin is to specifically facilitate formin-mediated actin assembly for cytokinesis in fission yeast.

Original languageEnglish (US)
Pages (from-to)283-293
Number of pages11
JournalMolecular biology of the cell
Volume26
Issue number2
DOIs
StatePublished - Jan 15 2015

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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