Structural pathway for PI3-kinase regulation by VPS15 in autophagy

Output Details

Preprint October 2, 2023

Published February 5, 2025

The class III phosphatidylinositol-3 kinase complexes I and II (PI3KC3-C1 and -C2) have vital roles in macroautophagy and endosomal maturation, respectively. We elucidated a structural pathway of enzyme activation through cryo-EM analysis of PI3KC3-C1. The inactive conformation of the VPS15 pseudokinase stabilizes the inactive conformation, sequestering its *N*-myristate in the N-lobe of the pseudokinase. Upon activation, the myristate is liberated such that the VPS34 lipid kinase catalyzes PI3P production on membranes. The VPS15 pseudokinase domain binds tightly to guanosine triphosphate (GTP), and stabilizes a web of interactions to autoinhibit the cytosolic complex and to promote the activation upon membrane binding. These findings show in atomistic detail how the VPS34 lipid kinase is activated in the context of a complete PI3K complex.
Identifier (DOI)
10.1126/science.adl3787
Tags
  • Cell biology
  • Molecular Dynamics simulation (MD)
  • Original Research
  • PI3KC3-C1 complex
  • Rab GTPases
  • Recombinant
  • Structural biology

Meet the Authors

  • Annan (Zeke) Cook, BSc

    Key Personnel: Team Hurley

    University of California, Berkeley

  • Minghao Chen, PhD

    Key Personnel: Team Hurley

    University of California, Berkeley

  • Thanh Nguyen, PhD

    Key Personnel: Team Hurley

    Walter and Eliza Hall Institute of Medical Research

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    Ainara Claveras Cabezudo, MSc

    Key Personnel: Team Hurley

    Max Planck Institute of Biophysics

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    Grace Khuu, BSc

    Key Personnel: Team Hurley

    Monash University

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    Shanlin Rao, PhD

    Key Personnel: Team Hurley

    Max Planck Institute of Biophysics

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    Samantha N. Garcia

  • User avatar fallback logo

    Mingxuan Yang

  • User avatar fallback logo

    Anthony T. Iavarone

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    Xuefeng Ren, PhD

    Key Personnel: Team Hurley

    University of California, Berkeley

  • Michael Lazarou, PhD

    Co-PI (Core Leadership): Team Hurley

    Monash University

  • Gerhard Hummer, PhD

    Collaborating PI: Team Hurley

    Max Planck Institute of Biophysics

  • James Hurley

    Lead PI (Core Leadership): Team Hurley

    University of California, Berkeley