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PMID: 26139537 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Crosstalk between Akt/GSK3β signaling and dynamin-1 regulates clathrin-mediated endocytosis.

The EMBO journal ·Vol. 34 ·No. 16 ·2015-08-13 ·Pages 2132-46

Reis CR, Chen PH, Srinivasan S, Aguet F, Mettlen M, Schmid SL

Abstract

Clathrin-mediated endocytosis (CME) regulates signaling from the plasma membrane. Analysis of clathrin-coated pit (CCP) dynamics led us to propose the existence of a rate-limiting, regulatory step(s) that monitor the fidelity of early stages in CCP maturation. Here we show that nascent endocytic vesicles formed in mutant cells displaying rapid, dysregulated CME are defective in early endosomal trafficking, maturation and acidification, confirming the importance of this "checkpoint." Dysregulated CME also alters EGF receptor signaling and leads to constitutive activation of the protein kinase Akt. Dynamin-1, which was thought to be neuron specific, is activated by the Akt/GSK3β signaling cascade in non-neuronal cells to trigger rapid, dysregulated CME. Acute activation of dynamin-1 in RPE cells by inhibition of GSK3β accelerates CME, alters CCP dynamics and, unexpectedly, increases the rate of CCP initiation. CRISPR-Cas9n-mediated knockout and reconstitution studies establish that dynamin-1 is activated by Akt/GSK3β signaling in H1299 non-small lung cancer cells. These findings provide direct evidence for an isoform-specific role for dynamin in regulating CME and reveal a feed-forward pathway that could link signaling from cell surface receptors to the regulation of CME.

Keywords
CCP maturation CME dynamin endosomal trafficking signaling
MeSH Terms
Cell Line Clathrin-Coated Vesicles/metabolism Dynamin I/metabolism Endocytosis Epithelial Cells/physiology Glycogen Synthase Kinase 3/metabolism Glycogen Synthase Kinase 3 beta Humans Protein Interaction Maps Proto-Oncogene Proteins c-akt/metabolism Signal Transduction
Chemicals
GSK3B protein, human Glycogen Synthase Kinase 3 beta Proto-Oncogene Proteins c-akt Glycogen Synthase Kinase 3 Dynamin I
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Reis Carlos R
Department of Cell Biology, UT Southwestern Medical Center, Dallas, TX, USA.
Chen Ping-Hung
Department of Cell Biology, UT Southwestern Medical Center, Dallas, TX, USA.
Srinivasan Saipraveen
Department of Cell Biology, UT Southwestern Medical Center, Dallas, TX, USA.
Aguet François
Department of Cell Biology, Harvard Medical School, Boston, MA, USA.
Mettlen Marcel
Department of Cell Biology, UT Southwestern Medical Center, Dallas, TX, USA.
Schmid Sandra L
Department of Cell Biology, UT Southwestern Medical Center, Dallas, TX, USA Sandra.Schmid@utsouthwestern.edu.
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.15252/embj.201591518
Email
sandra.schmid@utsouthwestern.edu
Published
2015-08-13
Epub
2015-00-02
Pages
2132-46
Language
English
Region
England
NLM ID
8208664
PMCID
PMC4557666
Subset
IM
Grants
NIMH NIH HHS · R37 MH061345 · United States
NIMH NIH HHS · MH61345 · United States
NIMH NIH HHS · R01 MH061345 · United States
NIGMS NIH HHS · GM42455 · United States
NIGMS NIH HHS · R01 GM73165 · United States
NIGMS NIH HHS · R01 GM042455 · United States
NIGMS NIH HHS · R01 GM073165 · United States
External Links
PubMed source DOI: 10.15252/embj.201591518 PMC Full Text View journal details
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