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Drosophila Cappuccino alleles provide insight into formin mechanism and role in oogenesis

During Drosophila development, the formin actin nucleator Cappuccino (Capu) helps build a cytoplasmic actin mesh throughout the oocyte. Loss of Capu leads to female sterility, presumably because polarity determinants fail to localize properly in the absence of the mesh. To gain deeper insight into h...

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Autores principales: Yoo, Haneul, Roth-Johnson, Elizabeth A., Bor, Batbileg, Quinlan, Margot E.
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/PMC4436832/
https://www.ncbi.nlm.nih.gov/pubmed/25788286
http://dx.doi.org/10.1091/mbc.E14-11-1558
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author Yoo, Haneul
Roth-Johnson, Elizabeth A.
Bor, Batbileg
Quinlan, Margot E.
author_facet Yoo, Haneul
Roth-Johnson, Elizabeth A.
Bor, Batbileg
Quinlan, Margot E.
author_sort Yoo, Haneul
collection PubMed
description During Drosophila development, the formin actin nucleator Cappuccino (Capu) helps build a cytoplasmic actin mesh throughout the oocyte. Loss of Capu leads to female sterility, presumably because polarity determinants fail to localize properly in the absence of the mesh. To gain deeper insight into how Capu builds this actin mesh, we systematically characterized seven capu alleles, which have missense mutations in Capu's formin homology 2 (FH2) domain. We report that all seven alleles have deleterious effects on fly fertility and the actin mesh in vivo but have strikingly different effects on Capu's biochemical activity in vitro. Using a combination of bulk and single- filament actin-assembly assays, we find that the alleles differentially affect Capu's ability to nucleate and processively elongate actin filaments. We also identify a unique “loop” in the lasso region of Capu's FH2 domain. Removing this loop enhances Capu's nucleation, elongation, and F-actin–bundling activities in vitro. Together our results on the loop and the seven missense mutations provides mechanistic insight into formin function in general and Capu's role in the Drosophila oocyte in particular.
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spelling pubmed-44368322015-07-30 Drosophila Cappuccino alleles provide insight into formin mechanism and role in oogenesis Yoo, Haneul Roth-Johnson, Elizabeth A. Bor, Batbileg Quinlan, Margot E. Mol Biol Cell Articles During Drosophila development, the formin actin nucleator Cappuccino (Capu) helps build a cytoplasmic actin mesh throughout the oocyte. Loss of Capu leads to female sterility, presumably because polarity determinants fail to localize properly in the absence of the mesh. To gain deeper insight into how Capu builds this actin mesh, we systematically characterized seven capu alleles, which have missense mutations in Capu's formin homology 2 (FH2) domain. We report that all seven alleles have deleterious effects on fly fertility and the actin mesh in vivo but have strikingly different effects on Capu's biochemical activity in vitro. Using a combination of bulk and single- filament actin-assembly assays, we find that the alleles differentially affect Capu's ability to nucleate and processively elongate actin filaments. We also identify a unique “loop” in the lasso region of Capu's FH2 domain. Removing this loop enhances Capu's nucleation, elongation, and F-actin–bundling activities in vitro. Together our results on the loop and the seven missense mutations provides mechanistic insight into formin function in general and Capu's role in the Drosophila oocyte in particular. The American Society for Cell Biology 2015-05-15 /pmc/articles/PMC4436832/ /pubmed/25788286 http://dx.doi.org/10.1091/mbc.E14-11-1558 Text en © 2015 Yoo, Roth-Johnson, 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
Yoo, Haneul
Roth-Johnson, Elizabeth A.
Bor, Batbileg
Quinlan, Margot E.
Drosophila Cappuccino alleles provide insight into formin mechanism and role in oogenesis
title Drosophila Cappuccino alleles provide insight into formin mechanism and role in oogenesis
title_full Drosophila Cappuccino alleles provide insight into formin mechanism and role in oogenesis
title_fullStr Drosophila Cappuccino alleles provide insight into formin mechanism and role in oogenesis
title_full_unstemmed Drosophila Cappuccino alleles provide insight into formin mechanism and role in oogenesis
title_short Drosophila Cappuccino alleles provide insight into formin mechanism and role in oogenesis
title_sort drosophila cappuccino alleles provide insight into formin mechanism and role in oogenesis
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4436832/
https://www.ncbi.nlm.nih.gov/pubmed/25788286
http://dx.doi.org/10.1091/mbc.E14-11-1558
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