The Actin Nucleator Cobl Is Controlled by Calcium and Calmodulin

PLoS Biology
Wenya HouBritta Qualmann

Abstract

Actin nucleation triggers the formation of new actin filaments and has the power to shape cells but requires tight control in order to bring about proper morphologies. The regulation of the members of the novel class of WASP Homology 2 (WH2) domain-based actin nucleators, however, thus far has largely remained elusive. Our study reveals signal cascades and mechanisms regulating Cordon-Bleu (Cobl). Cobl plays some, albeit not fully understood, role in early arborization of neurons and nucleates actin by a mechanism that requires a combination of all three of its actin monomer-binding WH2 domains. Our experiments reveal that Cobl is regulated by Ca2+ and multiple, direct associations of the Ca2+ sensor Calmodulin (CaM). Overexpression analyses and rescue experiments of Cobl loss-of-function phenotypes with Cobl mutants in primary neurons and in tissue slices demonstrated the importance of CaM binding for Cobl's functions. Cobl-induced dendritic branch initiation was preceded by Ca2+ signals and coincided with local F-actin and CaM accumulations. CaM inhibitor studies showed that Cobl-mediated branching is strictly dependent on CaM activity. Mechanistic studies revealed that Ca2+/CaM modulates Cobl's actin binding properties and f...Continue Reading

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Citations

Apr 1, 2017·Molecular and Cellular Neurosciences·Lorena Armijo-Weingart, Gianluca Gallo
Dec 14, 2017·The Journal of Cell Biology·Maryam IzadiMichael M Kessels
Sep 4, 2015·PLoS Biology·George Leondaritis, Britta Johanna Eickholt
Sep 25, 2019·Nature Cell Biology·David WolfBritta Qualmann
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Jun 3, 2017·Frontiers in Cellular Neuroscience·Anja KonietznyMarina Mikhaylova
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Methods Mentioned

BETA
Coprecipitation
immunoprecipitations
transfection
coimmunoprecipitations
coimmunoprecipitation
transfections
GTPases
PCR
immunoprecipitation

Software Mentioned

GraphPad Prism
LAS AF
ZEN2012
Adobe Illustrator
Adobe Photoshop
Imaris
AxioVision
ImageJ
ZEN
Excel

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