RT Journal Article
SR Electronic
T1 A new CRISPR screening approach for identifying novel autophagy-related factors and cytoplasm-to-lysosome trafficking routes
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 229732
DO 10.1101/229732
A1 Christopher J. Shoemaker
A1 Tina Q. Huang
A1 Nicholas R. Weir
A1 Nicole Polyakov
A1 Vladimir Denic
YR 2017
UL http://biorxiv.org/content/early/2017/12/06/229732.abstract
AB Selective autophagy comprises cytoplasm-to-lysosome trafficking routes that transport cargos using double-membrane vesicles (autophagosomes). Cargos are detected by receptor proteins, which typically also bind to lipid-conjugated LC3 proteins on autophagosome membranes. We dissected lysosomal delivery of four SQSTM1-like receptors by genome-wide CRISPR screening looking for novel autophagy-related (ATG) factors and trafficking routes. We uncovered new mammalian ATG factors including TMEM41B, an endoplasmic reticulum membrane protein required for autophagosome membrane expansion and/or closure. Furthermore, we found that certain receptors remain robustly targeted to the lysosome even in the absence of ATG7 or other LC3 conjugation factors. Lastly, we identified a unique genetic fingerprint behind receptor flux in ATG7KO cells, which includes factors implicated in nucleating autophagosome formation and vesicle trafficking factors not previously linked with autophagy. Our work uncovers new ATG factors, reveals a malleable network of autophagy receptor genetic interactions, and provides a valuable resource (http://35.196.197.20:3838/cscrispr/) for further mining of novel autophagy mechanisms.