Cell-substrate adhesion drives Scar/WAVE activation and phosphorylation by a Ste20-family kinase, which controls pseudopod lifetime.
Animals
CRISPR-Cas Systems
Cell Adhesion
Cell Line, Tumor
Chemotaxis
/ genetics
Dictyostelium
/ genetics
Gene Editing
/ methods
Gene Expression Regulation
MAP Kinase Kinase Kinase 3
/ genetics
Melanocytes
/ metabolism
Mice
Mitogen-Activated Protein Kinase 1
/ genetics
Mutation
NIH 3T3 Cells
Phenotype
Phosphorylation
Ploidies
Protozoan Proteins
/ genetics
Pseudopodia
/ genetics
Wiskott-Aldrich Syndrome Protein Family
/ genetics
Journal
PLoS biology
ISSN: 1545-7885
Titre abrégé: PLoS Biol
Pays: United States
ID NLM: 101183755
Informations de publication
Date de publication:
08 2020
08 2020
Historique:
received:
27
03
2020
accepted:
13
07
2020
revised:
13
08
2020
pubmed:
4
8
2020
medline:
18
9
2020
entrez:
4
8
2020
Statut:
epublish
Résumé
The Scar/WAVE complex is the principal catalyst of pseudopod and lamellipod formation. Here we show that Scar/WAVE's proline-rich domain is polyphosphorylated after the complex is activated. Blocking Scar/WAVE activation stops phosphorylation in both Dictyostelium and mammalian cells, implying that phosphorylation modulates pseudopods after they have been formed, rather than controlling whether they are initiated. Unexpectedly, phosphorylation is not promoted by chemotactic signaling but is greatly stimulated by cell:substrate adhesion and diminished when cells deadhere. Phosphorylation-deficient or phosphomimetic Scar/WAVE mutants are both normally functional and rescue the phenotype of knockout cells, demonstrating that phosphorylation is dispensable for activation and actin regulation. However, pseudopods and patches of phosphorylation-deficient Scar/WAVE last substantially longer in mutants, altering the dynamics and size of pseudopods and lamellipods and thus changing migration speed. Scar/WAVE phosphorylation does not require ERK2 in Dictyostelium or mammalian cells. However, the MAPKKK homologue SepA contributes substantially-sepA mutants have less steady-state phosphorylation, which does not increase in response to adhesion. The mutants also behave similarly to cells expressing phosphorylation-deficient Scar, with longer-lived pseudopods and patches of Scar recruitment. We conclude that pseudopod engagement with substratum is more important than extracellular signals at regulating Scar/WAVE's activity and that phosphorylation acts as a pseudopod timer by promoting Scar/WAVE turnover.
Identifiants
pubmed: 32745097
doi: 10.1371/journal.pbio.3000774
pii: PBIOLOGY-D-20-00795
pmc: PMC7425996
doi:
Substances chimiques
Protozoan Proteins
0
SCAR protein, Dictyostelium
0
Wiskott-Aldrich Syndrome Protein Family
0
Mitogen-Activated Protein Kinase 1
EC 2.7.11.24
MAP Kinase Kinase Kinase 3
EC 2.7.11.25
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e3000774Subventions
Organisme : Cancer Research UK
ID : A17196
Pays : United Kingdom
Organisme : Cancer Research UK
ID : A20017
Pays : United Kingdom
Organisme : Cancer Research UK
ID : A20017
Pays : United Kingdom
Organisme : NIGMS NIH HHS
ID : R35 GM134895
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM063691
Pays : United States
Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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