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PMID: 26450213 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Structure and flexibility of the endosomal Vps34 complex reveals the basis of its function on membranes.

Science (New York, N.Y.) ·Vol. 350 ·No. 6257 ·2015-10-09 ·Pages aac7365

Rostislavleva K, Soler N, Ohashi Y, Zhang L, Pardon E, Burke JE, Masson GR, Johnson C, Steyaert J, Ktistakis NT, Williams RL

Abstract

Phosphatidylinositol 3-kinase Vps34 complexes regulate intracellular membrane trafficking in endocytic sorting, cytokinesis, and autophagy. We present the 4.4 angstrom crystal structure of the 385-kilodalton endosomal complex II (PIK3C3-CII), consisting of Vps34, Vps15 (p150), Vps30/Atg6 (Beclin 1), and Vps38 (UVRAG). The subunits form a Y-shaped complex, centered on the Vps34 C2 domain. Vps34 and Vps15 intertwine in one arm, where the Vps15 kinase domain engages the Vps34 activation loop to regulate its activity. Vps30 and Vps38 form the other arm that brackets the Vps15/Vps34 heterodimer, suggesting a path for complex assembly. We used hydrogen-deuterium exchange mass spectrometry (HDX-MS) to reveal conformational changes accompanying membrane binding and identify a Vps30 loop that is critical for the ability of complex II to phosphorylate giant liposomes on which complex I is inactive.

MeSH Terms
Cell Membrane/chemistry,enzymology Class III Phosphatidylinositol 3-Kinases/chemistry,ultrastructure Crystallography, X-Ray Endosomes/chemistry,enzymology Protein Multimerization Protein Structure, Secondary Protein Structure, Tertiary Saccharomyces cerevisiae/enzymology Vacuolar Sorting Protein VPS15/chemistry,ultrastructure
Chemicals
Class III Phosphatidylinositol 3-Kinases Vacuolar Sorting Protein VPS15
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Rostislavleva Ksenia
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Soler Nicolas
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Ohashi Yohei
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Zhang Lufei
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Pardon Els
Structural Biology Research Center, VIB, B-1050 Brussels, Belgium. | Structural Biology Brussels, Vrije Universiteit Brussel, B-1050 Brussel, Belgium.
Burke John E
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Masson Glenn R
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Johnson Chris
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Steyaert Jan
Structural Biology Research Center, VIB, B-1050 Brussels, Belgium. | Structural Biology Brussels, Vrije Universiteit Brussel, B-1050 Brussel, Belgium.
Ktistakis Nicholas T
The Babraham Institute, Cambridge UK.
Williams Roger L
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
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Article Info
Journal
Science (New York, N.Y.)
Abbr.
Science
ISSN
1095-9203
DOI
10.1126/science.aac7365
Published
2015-10-09
Pages
aac7365
Language
English
Region
United States
NLM ID
0404511
PMCID
PMC4601532
Subset
IM
Grants
Biotechnology and Biological Sciences Research Council · BBS/E/B/00001221 · United Kingdom
British Heart Foundation · PG11/109/29247 · United Kingdom
Medical Research Council · U105184308 · United Kingdom
British Heart Foundation · PG/11/109/29247 · United Kingdom
Medical Research Council · MC_U105184308 · United Kingdom
Biotechnology and Biological Sciences Research Council · BB/K019155/1 · United Kingdom
Databases
PDB
5DFZ
External Links
PubMed source DOI: 10.1126/science.aac7365 PMC Full Text View journal details
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