Journal Article

An N-terminal conserved region in human Atg3 couples membrane curvature sensitivity to conjugase activity during autophagy

Flanagan, John M., Wang, Hong-Gang, Shen, Yan, Bui, Van, Tian, Fang, Ye, Yansheng, Jiang, Xuejun, Tyndall, Erin R., Tang, Zhenyuan

2021

Overview

During autophagy the enzyme Atg3 catalyzes the covalent conjugation of LC3 to the amino group of phosphatidylethanolamine (PE) lipids, which is one of the key steps in autophagosome formation. Here, we have demonstrated that an N-terminal conserved region of human Atg3 (hAtg3) communicates information from the N-terminal membrane curvature-sensitive amphipathic helix (AH), which presumably targets the enzyme to the tip of phagophore, to the C-terminally located catalytic core for LC3–PE conjugation. Mutations in the putative communication region greatly reduce or abolish the ability of hAtg3 to catalyze this conjugation in vitro and in vivo, and alter the membrane-bound conformation of the wild-type protein, as reported by NMR. Collectively, our results demonstrate that the N-terminal conserved region of hAtg3 works in concert with its geometry-selective AH to promote LC3–PE conjugation only on the target membrane, and substantiate the concept that highly curved membranes drive spatial regulation of the autophagosome biogenesis during autophagy. The E2-like enzyme human Atg3 catalyses the transfer of ubiquitin-like mammalian LC3 to the lipid phosphatidylethanolamine during autophagosome formation. Here, the authors combine NMR measurements with in vitro biochemical and in vivo cellular assays and show that the N-terminal conserved region of human Atg3 communicates information from the curvature-sensing domain to its active site.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 101/6 / 631/45/535/878/1263 / 631/45/607 / 631/57 / Autophagy / Autophagy-Related Proteins - chemistry / Autophagy-Related Proteins - genetics / Autophagy-Related Proteins - metabolism / Biocatalysis / Cell Membrane - genetics / Cell Membrane - metabolism / Conjugation / Curvature / Enzymes / Evolution / Humanities and Social Sciences / Humans / Lipids / Membranes / Microtubule-Associated Proteins - metabolism / multidisciplinary / Mutation / NMR / Nuclear magnetic resonance / Phagocytosis / Phosphatidylethanolamine / Protein Conformation, alpha-Helical / Protein Domains / Protein structure / Science / Science (multidisciplinary) / Ubiquitin / Ubiquitin-Conjugating Enzymes - chemistry / Ubiquitin-Conjugating Enzymes - genetics / Ubiquitin-Conjugating Enzymes - metabolism