Abstract
The class III phosphatidylinositol‐3‐kinase (PI3KC3) plays a central role in autophagy and endocytosis. How its function is differentially regulated in these processes remain elusive. We are the first group to purify PI3KC3 holocomplex in human cells, in which we found seven subunits including Vps34, p150, Beclin 1, Barkor/Atg14(L), NRBF2, Rubicon and UVRAG. These proteins form two mutually exclusive subcomplexes sharing Vps34, p150 and Beclin 1. Barkor and NRBF2 regulate PI3KC3 activity on autophagosome membrane, while UVRAG and Rubicon control PI3KC3 activity on endosome membrane. We have reconstituted the biochemical activity of autophagic PI3KC3 activity in vitro, and explored their functions in membrane curvature sensing and membrane tethering/hemifusion.
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