Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size.

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Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_01CBB053B8AE
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size.
Journal
eLife
Author(s)
Gerganova V., Floderer C., Archetti A., Michon L., Carlini L., Reichler T., Manley S., Martin S.G.
ISSN
2050-084X (Electronic)
ISSN-L
2050-084X
Publication state
Published
Issued date
03/05/2019
Peer-reviewed
Oui
Volume
8
Pages
e45983
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
Protein concentration gradients pattern developing organisms and single cells. In Schizosaccharomyces pombe rod-shaped cells, Pom1 kinase forms gradients with maxima at cell poles. Pom1 controls the timing of mitotic entry by inhibiting Cdr2, which forms stable membrane-associated nodes at mid-cell. Pom1 gradients rely on membrane association regulated by a phosphorylation-dephosphorylation cycle and lateral diffusion modulated by clustering. Using quantitative PALM imaging, we find individual Pom1 molecules bind the membrane too transiently to diffuse from pole to mid-cell. Instead, we propose they exchange within longer lived clusters forming the functional gradient unit. An allelic series blocking auto-phosphorylation shows that multi-phosphorylation shapes and buffers the gradient to control mid-cell levels, which represent the critical Cdr2-regulating pool. TIRF imaging of this cortical pool demonstrates more Pom1 overlaps with Cdr2 in short than long cells, consistent with Pom1 inhibition of Cdr2 decreasing with cell growth. Thus, the gradients modulate Pom1 mid-cell levels according to cell size.
Keywords
Cell Membrane/metabolism, Cytoplasm/enzymology, Phosphorylation, Protein Binding, Protein Kinases/analysis, Protein Kinases/metabolism, Protein Processing, Post-Translational, Protein-Serine-Threonine Kinases/metabolism, Schizosaccharomyces/enzymology, Schizosaccharomyces pombe Proteins/analysis, Schizosaccharomyces pombe Proteins/metabolism, Pom1 DYRK kinase, S. pombe, cell biology, cell size, concentration gradient, diffusion, single particle tracking PALM
Pubmed
Web of science
Open Access
Yes
Create date
06/05/2019 16:01
Last modification date
28/02/2020 6:19
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