Optogenetics reveals Cdc42 local activation by scaffold-mediated positive feedback and Ras GTPase.

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Version: Final published version
License: CC BY-NC-ND 4.0
Serval ID
serval:BIB_C2BCC5EE39F1
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Optogenetics reveals Cdc42 local activation by scaffold-mediated positive feedback and Ras GTPase.
Journal
PLoS biology
Author(s)
Lamas I., Merlini L., Vještica A., Vincenzetti V., Martin S.G.
ISSN
1545-7885 (Electronic)
ISSN-L
1544-9173
Publication state
Published
Issued date
01/2020
Peer-reviewed
Oui
Volume
18
Number
1
Pages
e3000600
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
Local activity of the small GTPase Cdc42 is critical for cell polarization. Whereas scaffold-mediated positive feedback was proposed to break symmetry of budding yeast cells and produce a single zone of Cdc42 activity, the existence of similar regulation has not been probed in other organisms. Here, we address this problem using rod-shaped cells of fission yeast Schizosaccharomyces pombe, which exhibit zones of active Cdc42-GTP at both cell poles. We implemented the CRY2-CIB1 optogenetic system for acute light-dependent protein recruitment to the plasma membrane, which allowed to directly demonstrate positive feedback. Indeed, optogenetic recruitment of constitutively active Cdc42 leads to co-recruitment of the guanine nucleotide exchange factor (GEF) Scd1 and endogenous Cdc42, in a manner dependent on the scaffold protein Scd2. We show that Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1). Remarkably, this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking. These cells further revealed that the small GTPase Ras1 plays a role in both localizing the GEF Scd1 and promoting its activity, which potentiates the positive feedback. We conclude that scaffold-mediated positive feedback, gated by Ras activity, confers robust polarization for rod-shape formation.
Keywords
Cell Polarity/genetics, Feedback, Physiological/physiology, Nuclear Matrix/physiology, Optogenetics, Organisms, Genetically Modified, Schizosaccharomyces/genetics, Schizosaccharomyces/metabolism, Schizosaccharomyces pombe Proteins/genetics, Schizosaccharomyces pombe Proteins/metabolism, cdc42 GTP-Binding Protein/genetics, cdc42 GTP-Binding Protein/metabolism, ras Proteins/physiology
Pubmed
Web of science
Open Access
Yes
Create date
29/01/2020 15:58
Last modification date
11/11/2020 8:10
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