Skip to Content
Merck
  • ATG8-Binding UIM Proteins Define a New Class of Autophagy Adaptors and Receptors.

ATG8-Binding UIM Proteins Define a New Class of Autophagy Adaptors and Receptors.

Cell (2019-04-09)
Richard S Marshall, Zhihua Hua, Sujina Mali, Fionn McLoughlin, Richard D Vierstra
ABSTRACT

During autophagy, vesicle dynamics and cargo recruitment are driven by numerous adaptors and receptors that become tethered to the phagophore through interactions with lipidated ATG8/LC3 decorating the expanding membrane. Most currently described ATG8-binding proteins exploit a well-defined ATG8-interacting motif (AIM, or LC3-interacting region [LIR]) that contacts a hydrophobic patch on ATG8 known as the LIR/AIM docking site (LDS). Here we describe a new class of ATG8 interactors that exploit ubiquitin-interacting motif (UIM)-like sequences for high-affinity binding to an alternative ATG8 interaction site. Assays with candidate UIM-containing proteins together with unbiased screens identified a large collection of UIM-based ATG8 interactors in plants, yeast, and humans. Analysis of a subset also harboring ubiquitin regulatory X (UBX) domains revealed a role for UIM-directed autophagy in clearing non-functional CDC48/p97 complexes, including some impaired in human disease. With this new class of adaptors and receptors, we greatly extend the reach of selective autophagy and identify new factors regulating autophagic vesicle dynamics.

MATERIALS
Product Number
Brand
Product Description

Millipore
BugBuster® Master Mix
Sigma-Aldrich
Sodium thiosulfate pentahydrate, ACS reagent, ≥99.5%
Sigma-Aldrich
Phenylmethanesulfonyl fluoride, ≥98.5% (GC)
Sigma-Aldrich
4-Nitrophenol, spectrophotometric grade
Sigma-Aldrich
2-Chloroacetamide, ≥98%
Millipore
Yeast Nitrogen Base Without Amino Acids and Ammonium Sulfate, suitable for microbiology, NutriSelect® Basic
Sigma-Aldrich
2-Chloroacetamide, ≥98.0% (HPLC)
Sigma-Aldrich
ML240, ≥98% (HPLC)
Sigma-Aldrich
Poly(ethylene glycol), average Mn 4,000, platelets
Sigma-Aldrich
3′,5′-Dimethoxy-4′-hydroxyacetophenone, 97%
Millipore
EZview Red Anti-HA Affinity Gel
Sigma-Aldrich
Lyticase from Arthrobacter luteus, lyophilized powder, ≥200 units/mg solid
Sigma-Aldrich
Gamborg′s B-5 Basal Medium with Minimal Organics, fine powder, suitable for plant cell culture
Sigma-Aldrich
Trifluoroacetic acid, suitable for HPLC, ≥99.0%