2. Academic Validation
  3. Vps34 Kinase Domain Dynamics Regulate the Autophagic PI 3-Kinase Complex

Vps34 Kinase Domain Dynamics Regulate the Autophagic PI 3-Kinase Complex

  • Mol Cell. 2017 Aug 3;67(3):528-534.e3. doi: 10.1016/j.molcel.2017.07.003.
Goran Stjepanovic 1,  Sulochanadevi Baskaran 2,  Mary G Lin 2,  James H Hurley 3
Affiliations

Affiliations

  • 1 Department of Molecular and Cell Biology and California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA 94720, USA; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
  • 2 Department of Molecular and Cell Biology and California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA 94720, USA.
  • 3 Department of Molecular and Cell Biology and California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA 94720, USA; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA. Electronic address: [email protected].
PMID: 28757208 DOI: 10.1016/j.molcel.2017.07.003
Abstract

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.

Keywords

allostery; autophagy; crosslinking mass spectrometry; electron microscopy; lipid kinase; phosphoinositide; protein dynamics; protein kinase; protein structure; pseudokinase.

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