The autophagy adaptor NDP52 and the FIP200 coiled-coil allosterically activate ULK1 complex membrane recruitment

Abstract
Data availability
Article and author information
Metrics

Abstract

The selective autophagy pathways of xenophagy and mitophagy are initiated when the adaptor NDP52 recruits the ULK1 complex to autophagic cargo. Hydrogen-deuterium exchange coupled to mass spectrometry (HDX-MS) was used to map the membrane and NDP52 binding sites of the ULK1 complex to unique regions of the coiled coil of the FIP200 subunit. Electron microscopy of the full-length ULK1 complex shows that the FIP200 coiled coil projects away from the crescent-shaped FIP200 N-terminal domain dimer. NDP52 allosterically stimulates membrane-binding by FIP200 and the ULK1 complex by promoting a more dynamic conformation of the membrane-binding portion of the FIP200 coiled coil. Giant unilamellar vesicle (GUV) reconstitution confirmed that membrane recruitment by the ULK1 complex is triggered by NDP52 engagement. These data reveal how the allosteric linkage between NDP52 and the ULK1 complex could drive the first membrane recruitment event of phagophore biogenesis in xenophagy and mitophagy.

Data availability

All data generated in this study are included in the manuscript and supporting files. HDX-MS data are included as Supplementary Data Set 1.

Article and author information

Author details

Xiaoshan Shi

Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-7931-8684
Chunmei Chang

Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

Competing interests
No competing interests declared.
Adam L Yokom

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0002-3746-7961
Liv E Jensen

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

Competing interests
No competing interests declared.
James H Hurley

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

For correspondence
jimhurley@berkeley.edu

Competing interests
James H Hurley, JHH is co-founder of Casma Therapeutics.

"This ORCID iD identifies the author of this article:" 0000-0001-5054-5445

Funding

National Institute of General Medical Sciences (R01 GM111730)

James H Hurley

Human Frontier Sciences Program (RPGO 2017/0026)

James H Hurley

Jane Coffin Childs Foundation

Adam L Yokom

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.