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The LC3-conjugation machinery specifies the loading of RNA-binding proteins into extracellular vesicles
Nature Cell Biology ( IF 17.728 ) Pub Date : 2020-01-13 , DOI: 10.1038/s41556-019-0450-y
Andrew M. Leidal; Hector H. Huang; Timothy Marsh; Tina Solvik; Dachuan Zhang; Jordan Ye; FuiBoon Kai; Juliet Goldsmith; Jennifer Y. Liu; Yu-Hsin Huang; Teresa Monkkonen; Ariadne Vlahakis; Eric J. Huang; Hani Goodarzi; Li Yu; Arun P. Wiita; Jayanta Debnath

Traditionally viewed as an autodigestive pathway, autophagy also facilitates cellular secretion; however, the mechanisms underlying these processes remain unclear. Here, we demonstrate that components of the autophagy machinery specify secretion within extracellular vesicles (EVs). Using a proximity-dependent biotinylation proteomics strategy, we identify 200 putative targets of LC3-dependent secretion. This secretome consists of a highly interconnected network enriched in RNA-binding proteins (RBPs) and EV cargoes. Proteomic and RNA profiling of EVs identifies diverse RBPs and small non-coding RNAs requiring the LC3-conjugation machinery for packaging and secretion. Focusing on two RBPs, heterogeneous nuclear ribonucleoprotein K (HNRNPK) and scaffold-attachment factor B (SAFB), we demonstrate that these proteins interact with LC3 and are secreted within EVs enriched with lipidated LC3. Furthermore, their secretion requires the LC3-conjugation machinery, neutral sphingomyelinase 2 (nSMase2) and LC3-dependent recruitment of factor associated with nSMase2 activity (FAN). Hence, the LC3-conjugation pathway controls EV cargo loading and secretion.
更新日期:2020-01-14

 

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