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

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 Sc...

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Autores principales: Bestul, Andrew J., Christensen, Jenna R., Grzegorzewska, Agnieszka P., Burke, Thomas A., Sees, Jennifer A., Carroll, Robert T., Sirotkin, Vladimir, Keenan, Robert J., Kovar, David R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4294675/
https://www.ncbi.nlm.nih.gov/pubmed/25392301
http://dx.doi.org/10.1091/mbc.E13-05-0281
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author Bestul, Andrew J.
Christensen, Jenna R.
Grzegorzewska, Agnieszka P.
Burke, Thomas A.
Sees, Jennifer A.
Carroll, Robert T.
Sirotkin, Vladimir
Keenan, Robert J.
Kovar, David R.
author_facet Bestul, Andrew J.
Christensen, Jenna R.
Grzegorzewska, Agnieszka P.
Burke, Thomas A.
Sees, Jennifer A.
Carroll, Robert T.
Sirotkin, Vladimir
Keenan, Robert J.
Kovar, David R.
author_sort Bestul, Andrew J.
collection PubMed
description 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.
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spelling pubmed-42946752015-03-30 Fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin Cdc12 Bestul, Andrew J. Christensen, Jenna R. Grzegorzewska, Agnieszka P. Burke, Thomas A. Sees, Jennifer A. Carroll, Robert T. Sirotkin, Vladimir Keenan, Robert J. Kovar, David R. Mol Biol Cell Articles 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. The American Society for Cell Biology 2015-01-15 /pmc/articles/PMC4294675/ /pubmed/25392301 http://dx.doi.org/10.1091/mbc.E13-05-0281 Text en © 2015 Bestul, Christensen, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
spellingShingle Articles
Bestul, Andrew J.
Christensen, Jenna R.
Grzegorzewska, Agnieszka P.
Burke, Thomas A.
Sees, Jennifer A.
Carroll, Robert T.
Sirotkin, Vladimir
Keenan, Robert J.
Kovar, David R.
Fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin Cdc12
title Fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin Cdc12
title_full Fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin Cdc12
title_fullStr Fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin Cdc12
title_full_unstemmed Fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin Cdc12
title_short Fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin Cdc12
title_sort fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin cdc12
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4294675/
https://www.ncbi.nlm.nih.gov/pubmed/25392301
http://dx.doi.org/10.1091/mbc.E13-05-0281
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