Saltar al contenido
Merck

ULK1 and ULK2 Regulate Stress Granule Disassembly Through Phosphorylation and Activation of VCP/p97.

Molecular cell (2019-04-14)
Bo Wang, Brian A Maxwell, Joung Hyuck Joo, Youngdae Gwon, James Messing, Ashutosh Mishra, Timothy I Shaw, Amber L Ward, Honghu Quan, Sadie Miki Sakurada, Shondra M Pruett-Miller, Tulio Bertorini, Peter Vogel, Hong Joo Kim, Junmin Peng, J Paul Taylor, Mondira Kundu
RESUMEN

Disturbances in autophagy and stress granule dynamics have been implicated as potential mechanisms underlying inclusion body myopathy (IBM) and related disorders. Yet the roles of core autophagy proteins in IBM and stress granule dynamics remain poorly characterized. Here, we demonstrate that disrupted expression of the core autophagy proteins ULK1 and ULK2 in mice causes a vacuolar myopathy with ubiquitin and TDP-43-positive inclusions; this myopathy is similar to that caused by VCP/p97 mutations, the most common cause of familial IBM. Mechanistically, we show that ULK1/2 localize to stress granules and phosphorylate VCP, thereby increasing VCP's activity and ability to disassemble stress granules. These data suggest that VCP dysregulation and defective stress granule disassembly contribute to IBM-like disease in Ulk1/2-deficient mice. In addition, stress granule disassembly is accelerated by an ULK1/2 agonist, suggesting ULK1/2 as targets for exploiting the higher-order regulation of stress granules for therapeutic intervention of IBM and related disorders.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Dimetilsulfóxido, for molecular biology
Sigma-Aldrich
ANTI-FLAG® M2 monoclonal antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
Sigma-Aldrich
Bafilomycin A1 from Streptomyces griseus, ≥90% (HPLC)
Sigma-Aldrich
Carbonyl cyanide 3-chlorophenylhydrazone, ≥97% (TLC), powder
Sigma-Aldrich
Sample Buffer, Laemmli 2× Concentrate
Sigma-Aldrich
Anti-GAPDH antibody produced in rabbit, ~1 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Rosetta(DE3) Competent Cells - Novagen, Rosetta host strains are BL21 derivatives designed to enhance the expression of eukaryotic proteins that contain codons rarely used in E. coli.
Roche
FastStart TaqMan® Probe Master, sufficient for 100 reactions, sufficient for 500 reactions, sufficient for 2000 reactions, suitable for qPCR, suitable for RT-qPCR
Sigma-Aldrich
SBI-0206965, ≥98% (HPLC)
Sigma-Aldrich
LYN-1604 hydrochloride, ≥98% (HPLC)
Sigma-Aldrich
MISSION® esiRNA, targeting human VCP
Sigma-Aldrich
MISSION® esiRNA, targeting human ULK2