Vps34 Kinase Domain Dynamics Regulate the Autophagic PI 3-Kinase Complex.
The class III phosphatidylinositol 3-kinase complex I (PI3KC3-C1) is required for the initiation of essentially all macroautophagic processes. PI3KC3-C1 consists of the lipid kinase catalytic subunit VPS34, the VPS15 scaffold, and the regulatory BECN1 and ATG14 subunits. The VPS34 catalytic domain and BECN1:ATG14 subcomplex do not touch, and it is ... unclear how allosteric signals are transmitted to VPS34. We used EM and crosslinking mass spectrometry to dissect five conformational substates of the complex, including one in which the VPS34 catalytic domain is dislodged from the complex but remains tethered by an intrinsically disordered linker. A "leashed" construct prevented dislodging without interfering with the other conformations, blocked enzyme activity in vitro, and blocked autophagy induction in yeast cells. This pinpoints the dislodging and tethering of the VPS34 catalytic domain, and its regulation by VPS15, as a master allosteric switch in autophagy induction.
Mesh Terms:
Adaptor Proteins, Vesicular Transport, Allosteric Regulation, Autophagy, Autophagy-Related Proteins, Beclin-1, Class III Phosphatidylinositol 3-Kinases, HEK293 Cells, Humans, Mass Spectrometry, Mutation, Protein Interaction Domains and Motifs, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Signal Transduction, Structure-Activity Relationship, Vacuolar Sorting Protein VPS15
Adaptor Proteins, Vesicular Transport, Allosteric Regulation, Autophagy, Autophagy-Related Proteins, Beclin-1, Class III Phosphatidylinositol 3-Kinases, HEK293 Cells, Humans, Mass Spectrometry, Mutation, Protein Interaction Domains and Motifs, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Signal Transduction, Structure-Activity Relationship, Vacuolar Sorting Protein VPS15
Mol. Cell
Date: Aug. 03, 2017
PubMed ID: 28757208
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