Premium
Fluorescence‐based ATG 8 sensors monitor localization and function of LC 3/ GABARAP proteins
Author(s) -
Stolz Alexandra,
Putyrski Mateusz,
Kutle Ivana,
Huber Jessica,
Wang Chunxin,
Major Viktória,
Sidhu Sachdev S,
Youle Richard J,
Rogov Vladimir V,
Dötsch Volker,
Ernst Andreas,
Dikic Ivan
Publication year - 2016
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.201695063
Subject(s) - biology , mitophagy , microbiology and biotechnology , autophagy , mitochondrion , organelle , biogenesis , biochemistry , gene , apoptosis
Autophagy is a cellular surveillance pathway that balances metabolic and energy resources and transports specific cargos, including damaged mitochondria, other broken organelles, or pathogens for degradation to the lysosome. Central components of autophagosomal biogenesis are six members of the LC 3 and GABARAP family of ubiquitin‐like proteins ( mATG 8s). We used phage display to isolate peptides that possess bona fide LIR ( LC 3‐interacting region) properties and are selective for individual mATG 8 isoforms. Sensitivity of the developed sensors was optimized by multiplication, charge distribution, and fusion with a membrane recruitment ( FYVE ) or an oligomerization ( PB 1) domain. We demonstrate the use of the engineered peptides as intracellular sensors that recognize specifically GABARAP , GABL 1, GABL 2, and LC 3C, as well as a bispecific sensor for LC 3A and LC 3B. By using an LC 3C‐specific sensor, we were able to monitor recruitment of endogenous LC 3C to Salmonella during xenophagy, as well as to mitochondria during mitophagy. The sensors are general tools to monitor the fate of mATG 8s and will be valuable in decoding the biological functions of the individual LC 3/GABARAPs.