Endomembrane systems are reorganized by ORF3a and Membrane (M) of SARS-CoV-2 [article]

Yun-Bin Lee, Minkyo Jung, Jeesoo Kim, Myeong-Gyun Kang, Chulhwan Kwak, Jong-Seo Kim, Ji-Young Mun, Hyun-Woo Rhee
2021 bioRxiv   pre-print
The endomembrane reticulum (ER) is largely reorganized by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-CoV-2 ORF3a and membrane (M) protein expression affects ER-derived structures including cubic membrane and double membrane vesicles in coronavirus-infected cells; however, the molecular mechanisms underlying ER remodeling remain unclear. We introduced a 'plug and playable' proximity labeling tool (TurboID-GBP) for interactome mapping of GFP-tagged SARS-CoV-2 ORF3a and M
more » ... oteins. Through mass spectrometric identification of the biotinylated lysine residue (K+226 Da) on the viral proteins using Spot-TurboID workflow, 117 and 191 proteins were robustly determined as ORF3a and M interactomes, respectively, and many, including RNF5 (E3 ubiquitin ligase), overlap with the mitochondrial-associated membrane (MAM) proteome. RNF5 expression was correlated to ORF3a ubiquitination. MAM formation and secreted proteome profiles were largely affected by ORF3a expression. Thus, SARS-CoV-2 may utilize MAM as a viral assembly site, suggesting novel anti-viral treatment strategies for blocking viral replication in host cells.
doi:10.1101/2021.06.01.446555 fatcat:swzssx5fzvgy5aebyuwnxp456i