YWHAE/14-3-3ε crotonylation regulates leucine deprivation-induced autophagy

Zilong Zheng; Qing Zhong; Xianghua Yan

doi:10.1080/15548627.2023.2166276

YWHAE/14-3-3ε crotonylation regulates leucine deprivation-induced autophagy

Autophagy. 2023 Aug;19(8):2401-2402. doi: 10.1080/15548627.2023.2166276. Epub 2023 Jan 16.

Authors

Zilong Zheng^{1

2

3}, Qing Zhong⁴, Xianghua Yan^{1

2

3}

Affiliations

¹ State Key Laboratory of Agricultural Microbiology, Hubei Hongshan Laboratory, Frontiers Science Center for Animal Breeding and Sustainable Production, College of Animal Sciences and Technology, Huazhong Agricultural University, Wuhan, Hubei, China.
² The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan, Hubei, China.
³ Hubei Provincial Engineering Laboratory for Pig Precision Feeding and Feed Safety Technology, Huazhong Agricultural University, Wuhan, Hubei, China.
⁴ Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Department of Pathophysiology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.

Abstract

Macroautophagy/autophagy is an important process responsible for protein turnover and cell survival in amino acid-deprived conditions, especially for leucine (Leu). With the dramatic advances in mass spectrometry, many new post-translational modifications (PTMs) have been identified. However, whether these PTMs regulate autophagy remains unclear. Here we found global lysine crotonylation levels are significantly upregulated during Leu deprivation-induced autophagy. A comprehensive crotonylome profiling showed that YWHA/14-3-3 proteins are significantly enriched in the Leu regulated-crotonylome. The inhibition of YWHAE/14-3-3ε crotonylation by mutating two crotonylated sites to arginine, K73R K78R, significantly attenuates autophagy induced by Leu deprivation. Molecular dynamics suggest that YWHAE K73 and K78 crotonylations decrease protein conformation and thermodynamic stability. Moreover, we found crotonylation of YWHAE releases PPM1B to dephosphorylate ULK1 and consequently activate autophagy. Decrotonylation of YWHAE is mediated by HDAC7 whose activity is inhibited significantly by Leu deprivation. Taken together, our finding reveals a critical role of YWHAE crotonylation in Leu deprivation-induced autophagy.

Keywords: 14-3-3ε; HDAC7; PPM1B; autophagy; crotonylation; leucine deprivation.

Publication types

Research Support, Non-U.S. Gov't
Comment

MeSH terms

14-3-3 Proteins*
Autophagy*
Leucine / pharmacology
Mass Spectrometry
Protein Processing, Post-Translational

Substances

Leucine
14-3-3 Proteins

Grants and funding

This study was supported by the National Natural Science Foundation of China (31925037, 31520103915), the National Key Basic Research Program of China (973 Program) (2013CB127305).