Identification of CUL4A-DDB1-WDFY1 as an E3 ubiquitin ligase complex involved in initiation of lysophagy

Hirofumi Teranishi; Keisuke Tabata; Marika Saeki; Tetsuo Umemoto; Tomohisa Hatta; Takanobu Otomo; Kentaro Yamamoto; Toru Natsume; Tamotsu Yoshimori; Maho Hamasaki

doi:10.1016/j.celrep.2022.111349

Identification of CUL4A-DDB1-WDFY1 as an E3 ubiquitin ligase complex involved in initiation of lysophagy

Cell Rep. 2022 Sep 13;40(11):111349. doi: 10.1016/j.celrep.2022.111349.

Authors

Affiliations

¹ JT Pharmaceutical Frontier Research Laboratory, Yokohama 236-0004, Japan.
² Laboratory of Intracellular Membrane Dynamics, Graduate School of Frontier Biosciences Osaka University, Osaka 565-0871, Japan; Department of Genetics, Graduate School of Medicine Osaka University, Osaka 565-0871, Japan.
³ Laboratory of Intracellular Membrane Dynamics, Graduate School of Frontier Biosciences Osaka University, Osaka 565-0871, Japan.
⁴ Molecular Profiling Research Center for Drug Discovery, AIST, Tokyo 135-0064, Japan.
⁵ Department of Genetics, Graduate School of Medicine Osaka University, Osaka 565-0871, Japan.
⁶ Laboratory of Intracellular Membrane Dynamics, Graduate School of Frontier Biosciences Osaka University, Osaka 565-0871, Japan; Department of Genetics, Graduate School of Medicine Osaka University, Osaka 565-0871, Japan; Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University, Osaka 565-0871, Japan. Electronic address: tamyoshi@fbs.osaka-u.ac.jp.
⁷ Laboratory of Intracellular Membrane Dynamics, Graduate School of Frontier Biosciences Osaka University, Osaka 565-0871, Japan; Department of Genetics, Graduate School of Medicine Osaka University, Osaka 565-0871, Japan. Electronic address: hamasaki@fbs.osaka-u.ac.jp.

PMID: 36103833
DOI: 10.1016/j.celrep.2022.111349

Abstract

Macroautophagy is a bulk degradation system in which double membrane-bound structures called autophagosomes to deliver cytosolic materials to lysosomes. Autophagy promotes cellular homeostasis by selectively recognizing and sequestering specific targets, such as damaged organelles, protein aggregates, and invading bacteria, termed selective autophagy. We previously reported a type of selective autophagy, lysophagy, which helps clear damaged lysosomes. Damaged lysosomes become ubiquitinated and recruit autophagic machinery. Proteomic studies using transfection reagent-coated beads and further evaluations reveal that a CUL4A-DDB1-WDFY1 E3 ubiquitin ligase complex is essential to initiate lysophagy and clear damaged lysosomes. Moreover, we show that LAMP2 is ubiquitinated by the CUL4A E3 ligase complex as a substrate on damaged lysosomes. These results reveal how cells selectively tag damaged lysosomes to initiate autophagy for the clearance of lysosomes.

Keywords: CP: Molecular biology; CUL4A; LAMP2; autophagy; lysophagy; lysosomal membrane damage; ubiquitination.

Publication types

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

MeSH terms

Lysosomes / metabolism
Macroautophagy*
Proteomics*
Ubiquitin / metabolism
Ubiquitin-Protein Ligases / metabolism

Substances

Ubiquitin
Ubiquitin-Protein Ligases