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ULK1 induces autophagy by phosphorylating Beclin-1 and activating VPS34 lipid kinase

Nature Cell Biology volume 15, pages 741–750 (2013)Cite this article

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Abstract

Autophagy is the primary cellular catabolic program activated in response to nutrient starvation. Initiation of autophagy, particularly by amino-acid withdrawal, requires the ULK kinases. Despite its pivotal role in autophagy initiation, little is known about the mechanisms by which ULK promotes autophagy. Here we describe a molecular mechanism linking ULK to the pro-autophagic lipid kinase VPS34. Following amino-acid starvation or mTOR inhibition, the activated ULK1 phosphorylates Beclin-1 on Ser 14, thereby enhancing the activity of the ATG14L-containing VPS34 complexes. The Beclin-1 Ser 14 phosphorylation by ULK is required for full autophagic induction in mammals and this requirement is conserved in Caenorhabditis elegans. Our study reveals a molecular link from ULK1 to activation of the autophagy-specific VPS34 complex and autophagy induction.

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Figure 1: ULK is essential for activation of the ATG14L-associated VPS34 following amino-acid starvation.
Figure 2: Beclin-1 Ser 14 is phosphorylated by ULK1 and required for VPS34 activation in response to amino-acid withdrawal.
Figure 3: Beclin-1 is a physiological target of ULK kinase in response to amino-acid withdrawal and mTOR inhibition.
Figure 4: ATG14L stimulates Beclin-1 Ser 14 phosphorylation by promoting association with ULK1.
Figure 5: UVRAG promotes Beclin-1 Ser 14 phosphorylation and association with ULK1.
Figure 6: Beclin-1 Ser 14 phosphorylation plays a critical role in autophagy induction by amino-acid starvation.
Figure 7: The conserved ULK phosphorylation site in C. elegans Bec-1 is required for autophagy.
Figure 8: A working model of VPS34 complex regulation by ULK on amino-acid withdrawal.

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Acknowledgements

We would like to thank H. Zhang for LGG-1 and PGL-1 antibodies; J. Jewell, C. Hansen and K. Tumaneng for critical reading of this manuscript; and M. Farquhar for electron microscopy. Phosphosite identification by mass spectrometry was performed by the Proteomics Facility at the Fred Hutchinson Cancer Research Center. Confocal analysis was performed at the UCSD Neuroscience Microscopy Shared Facility (Grant P30 NS047101). This work was supported by National Institutes of Health (NIH) grants GM51586, GM62694 and CA108941, and the Department of Defense (W81XWH-0901-0279). R.C.R. is supported by a Canadian Institutes of Health Research (CIHR) postdoctoral fellowship.

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Authors and Affiliations

  1. Department of Pharmacology and Moores Cancer Center, University of California San Diego, La Jolla, California 92093, USA

    Ryan C. Russell, Haixin Yuan, Hyun Woo Park, Joungmok Kim, Haerin Kim & Kun-Liang Guan

  2. Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California 94720, USA

    Ye Tian & Andrew Dillin

  3. Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455, USA

    Yu-Yun Chang & Thomas P. Neufeld

  4. Department of Oral Biochemistry, School of Dentistry, Kyung Hee University, Seoul 130-701, Korea

    Joungmok Kim

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Contributions

R.C.R. planned and performed experiments, Y.T. planned and performed experiments, H.Y., H.W.P., Y-Y.C and H.K. performed experiments, J.K., T.P.N., A.D. and K-L.G. planned experiments, R.C.R. and K-L.G. wrote the manuscript.

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Correspondence to Kun-Liang Guan.

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Russell, R., Tian, Y., Yuan, H. et al. ULK1 induces autophagy by phosphorylating Beclin-1 and activating VPS34 lipid kinase. Nat Cell Biol 15, 741–750 (2013). https://doi.org/10.1038/ncb2757

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