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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2015
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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. |
format | Online Article Text |
id | pubmed-4436832 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
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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