Structural and functional analysis of the GABARAP interaction motif (GIM)
Abstract
Through the canonical LC3 interaction motif (LIR), [W/F/Y]‐X1‐X2‐[I/L/V], protein complexes are recruited to autophagosomes to perform their functions as either autophagy adaptors or receptors. How these adaptors/receptors selectively interact with either LC3 or GABARAP families remains unclear. Herein, we determine the range of selectivity of 30 known core LIR motifs towards individual LC3s and GABARAPs. From these, we define a GABARAP Interaction Motif (GIM) sequence ([W/F]‐[V/I]‐X2‐V) that the adaptor protein PLEKHM1 tightly conforms to. Using biophysical and structural approaches, we show that the PLEKHM1‐LIR is indeed 11‐fold more specific for GABARAP than LC3B. Selective mutation of the X1 and X2 positions either completely abolished the interaction with all LC3 and GABARAPs or increased PLEKHM1‐GIM selectivity 20‐fold towards LC3B. Finally, we show that conversion of p62/SQSTM1, FUNDC1 and FIP200 LIRs into our newly defined GIM, by introducing two valine residues, enhances their interaction with endogenous GABARAP over LC3B. The identification of a GABARAP‐specific interaction motif will aid the identification and characterization of the expanding array of autophagy receptor and adaptor proteins and their in vivo functions.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jun 27, 2017
- Source ID
- 10.15252/embr.201643587
Entities
People
- Alexandra Stolz
- Andreas Kniss
- Arvind C Ravichandran
- David G McEwan
- Diana O Rios‐szwed
- Frank Löhr
- Hironori Suzuki
- Ivan Đikić
- Renwick Dobson
- Soichi Wakatsuki
- Vladimir V Rogov
- Volker Dötsch
Organizations
- Army Research Office
- Fritz Thyssen Foundation
- German Research Foundation
- Goethe University Frankfurt
- Hessian Ministry for Science and the Arts
- Human Frontier Science Program
- Japan Society for the Promotion of Science
- Ministry of Business, Innovation and Employment
- Royal Society Te Apārangi
- Tenovus
- United States Army Research Laboratory
- University of Canterbury
- University of Dundee
- University of Melbourne
- Wellcome Trust