Recapitulating Actin Module Organization in the <i>Drosophila</i> Oocyte Reveals New Roles for Bristle-Actin-Modulating Proteins
The generation of F-actin bundles is controlled by the action of actin-binding proteins. In <i>Drosophila</i> bristle development, two major actin-bundling proteins—Forked and Fascin—were identified, but still the molecular mechanism by which these actin-bundling proteins and other prote...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2021-04-01
|
Series: | International Journal of Molecular Sciences |
Subjects: | |
Online Access: | https://www.mdpi.com/1422-0067/22/8/4006 |
id |
doaj-20e97f18cf9749e587408e6467bc8a44 |
---|---|
record_format |
Article |
spelling |
doaj-20e97f18cf9749e587408e6467bc8a442021-04-13T23:03:52ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-04-01224006400610.3390/ijms22084006Recapitulating Actin Module Organization in the <i>Drosophila</i> Oocyte Reveals New Roles for Bristle-Actin-Modulating ProteinsRamesh Kumar Krishnan0Raju Baskar1Bakhrat Anna2Natalie Elia3Mandy Boermel4Andreas R. Bausch5Uri Abdu6Department of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva 84105, IsraelDepartment of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva 84105, IsraelDepartment of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva 84105, IsraelDepartment of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva 84105, IsraelElectron Microscopy Core Facility, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117 Heidelberg, GermanyLehrstuhl für Zellbiophysik E27, Technische Universität München, James-Franck-Str. 1, 85748 Garching, GermanyDepartment of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva 84105, IsraelThe generation of F-actin bundles is controlled by the action of actin-binding proteins. In <i>Drosophila</i> bristle development, two major actin-bundling proteins—Forked and Fascin—were identified, but still the molecular mechanism by which these actin-bundling proteins and other proteins generate bristle actin bundles is unknown. In this study, we developed a technique that allows recapitulation of bristle actin module organization using the <i>Drosophila</i> ovary by a combination of confocal microscopy, super-resolution structured illumination microscopy, and correlative light and electron microscope analysis. Since Forked generated a distinct ectopic network of actin bundles in the oocyte, the additive effect of two other actin-associated proteins, namely, Fascin and Javelin (Jv), was studied. We found that co-expression of Fascin and Forked demonstrated that the number of actin filaments within the actin bundles dramatically increased, and in their geometric organization, they resembled bristle-like actin bundles. On the other hand, co-expression of Jv with Forked increased the length and density of the actin bundles. When all three proteins co-expressed, the actin bundles were longer and denser, and contained a high number of actin filaments in the bundle. Thus, our results demonstrate that the <i>Drosophila</i> oocyte could serve as a test tube for actin bundle analysis.https://www.mdpi.com/1422-0067/22/8/4006actin bundlesbristle<i>Drosophila</i>Fascinoocyte |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ramesh Kumar Krishnan Raju Baskar Bakhrat Anna Natalie Elia Mandy Boermel Andreas R. Bausch Uri Abdu |
spellingShingle |
Ramesh Kumar Krishnan Raju Baskar Bakhrat Anna Natalie Elia Mandy Boermel Andreas R. Bausch Uri Abdu Recapitulating Actin Module Organization in the <i>Drosophila</i> Oocyte Reveals New Roles for Bristle-Actin-Modulating Proteins International Journal of Molecular Sciences actin bundles bristle <i>Drosophila</i> Fascin oocyte |
author_facet |
Ramesh Kumar Krishnan Raju Baskar Bakhrat Anna Natalie Elia Mandy Boermel Andreas R. Bausch Uri Abdu |
author_sort |
Ramesh Kumar Krishnan |
title |
Recapitulating Actin Module Organization in the <i>Drosophila</i> Oocyte Reveals New Roles for Bristle-Actin-Modulating Proteins |
title_short |
Recapitulating Actin Module Organization in the <i>Drosophila</i> Oocyte Reveals New Roles for Bristle-Actin-Modulating Proteins |
title_full |
Recapitulating Actin Module Organization in the <i>Drosophila</i> Oocyte Reveals New Roles for Bristle-Actin-Modulating Proteins |
title_fullStr |
Recapitulating Actin Module Organization in the <i>Drosophila</i> Oocyte Reveals New Roles for Bristle-Actin-Modulating Proteins |
title_full_unstemmed |
Recapitulating Actin Module Organization in the <i>Drosophila</i> Oocyte Reveals New Roles for Bristle-Actin-Modulating Proteins |
title_sort |
recapitulating actin module organization in the <i>drosophila</i> oocyte reveals new roles for bristle-actin-modulating proteins |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-04-01 |
description |
The generation of F-actin bundles is controlled by the action of actin-binding proteins. In <i>Drosophila</i> bristle development, two major actin-bundling proteins—Forked and Fascin—were identified, but still the molecular mechanism by which these actin-bundling proteins and other proteins generate bristle actin bundles is unknown. In this study, we developed a technique that allows recapitulation of bristle actin module organization using the <i>Drosophila</i> ovary by a combination of confocal microscopy, super-resolution structured illumination microscopy, and correlative light and electron microscope analysis. Since Forked generated a distinct ectopic network of actin bundles in the oocyte, the additive effect of two other actin-associated proteins, namely, Fascin and Javelin (Jv), was studied. We found that co-expression of Fascin and Forked demonstrated that the number of actin filaments within the actin bundles dramatically increased, and in their geometric organization, they resembled bristle-like actin bundles. On the other hand, co-expression of Jv with Forked increased the length and density of the actin bundles. When all three proteins co-expressed, the actin bundles were longer and denser, and contained a high number of actin filaments in the bundle. Thus, our results demonstrate that the <i>Drosophila</i> oocyte could serve as a test tube for actin bundle analysis. |
topic |
actin bundles bristle <i>Drosophila</i> Fascin oocyte |
url |
https://www.mdpi.com/1422-0067/22/8/4006 |
work_keys_str_mv |
AT rameshkumarkrishnan recapitulatingactinmoduleorganizationintheidrosophilaioocyterevealsnewrolesforbristleactinmodulatingproteins AT rajubaskar recapitulatingactinmoduleorganizationintheidrosophilaioocyterevealsnewrolesforbristleactinmodulatingproteins AT bakhratanna recapitulatingactinmoduleorganizationintheidrosophilaioocyterevealsnewrolesforbristleactinmodulatingproteins AT natalieelia recapitulatingactinmoduleorganizationintheidrosophilaioocyterevealsnewrolesforbristleactinmodulatingproteins AT mandyboermel recapitulatingactinmoduleorganizationintheidrosophilaioocyterevealsnewrolesforbristleactinmodulatingproteins AT andreasrbausch recapitulatingactinmoduleorganizationintheidrosophilaioocyterevealsnewrolesforbristleactinmodulatingproteins AT uriabdu recapitulatingactinmoduleorganizationintheidrosophilaioocyterevealsnewrolesforbristleactinmodulatingproteins |
_version_ |
1721528298188570624 |