Architecture and dynamics of the autophagic phosphatidylinositol 3-kinase complex
Open Access
- 9 December 2014
- journal article
- research article
- Published by eLife Sciences Publications, Ltd in eLife
Abstract
The class III phosphatidylinositol 3-kinase complex I (PI3KC3-C1) that functions in early autophagy consists of the lipid kinase VPS34, the scaffolding protein VPS15, the tumor suppressor BECN1, and the autophagy-specific subunit ATG14. The structure of the ATG14-containing PI3KC3-C1 was determined by single-particle EM, revealing a V-shaped architecture. All of the ordered domains of VPS34, VPS15, and BECN1 were mapped by MBP tagging. The dynamics of the complex were defined using hydrogen–deuterium exchange, revealing a novel 20-residue ordered region C-terminal to the VPS34 C2 domain. VPS15 organizes the complex and serves as a bridge between VPS34 and the ATG14:BECN1 subcomplex. Dynamic transitions occur in which the lipid kinase domain is ejected from the complex and VPS15 pivots at the base of the V. The N-terminus of BECN1, the target for signaling inputs, resides near the pivot point. These observations provide a framework for understanding the allosteric regulation of lipid kinase activity.Keywords
Funding Information
- National Institute of General Medical Sciences (GM111730)
- Howard Hughes Medical Institute (HHMI)
- Damon Runyon Cancer Research Foundation (Fellowship)
- L'Oreal USA (Women in Science Fellowship)
- Human Frontier Science Program (Long-Term Fellowship (LT001037/2011-L))
This publication has 66 references indexed in Scilit:
- Differential Regulation of Distinct Vps34 Complexes by AMPK in Nutrient Stress and AutophagyCell, 2013
- Mechanisms of Autophagosome BiogenesisCurrent Biology, 2012
- Methods for the analysis of high precision differential hydrogen–deuterium exchange dataInternational Journal of Mass Spectrometry, 2011
- The Beclin 1 interactomeCurrent Opinion in Cell Biology, 2010
- Hydrogen exchange and mass spectrometry: A historical perspectiveJournal of the American Society for Mass Spectrometry, 2006
- Automated molecular microscopy: The new Leginon systemJournal of Structural Biology, 2005
- UCSF Chimera—A visualization system for exploratory research and analysisJournal of Computational Chemistry, 2004
- EMAN: Semiautomated Software for High-Resolution Single-Particle ReconstructionsJournal of Structural Biology, 1999
- SPIDER and WEB: Processing and Visualization of Images in 3D Electron Microscopy and Related FieldsJournal of Structural Biology, 1996
- A New Generation of the IMAGIC Image Processing SystemJournal of Structural Biology, 1996