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Discovery and functional interrogation of SARS-CoV-2 protein-RNA interactions #MMPMID35233578
Xiang JS; Mueller JR; Luo EC; Yee BA; Schafer D; Schmok JC; Tan FE; Rothamel K; McVicar RN; Kwong EM; Jones KL; Her HL; Chen CY; Vu AQ; Jin W; Park SS; Le P; Brannan KW; Kofman ER; Li Y; Tankka AT; Dong KD; Song Y; Carlin AF; Van Nostrand EL; Leibel SL; Yeo GW
bioRxiv 2022[Feb]; ä (ä): ä PMID35233578show ga
The COVID-19 pandemic is caused by severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2). The betacoronvirus has a positive sense RNA genome which encodes for several RNA binding proteins. Here, we use enhanced crosslinking and immunoprecipitation to investigate SARS-CoV-2 protein interactions with viral and host RNAs in authentic virus-infected cells. SARS-CoV-2 proteins, NSP8, NSP12, and nucleocapsid display distinct preferences to specific regions in the RNA viral genome, providing evidence for their shared and separate roles in replication, transcription, and viral packaging. SARS-CoV-2 proteins expressed in human lung epithelial cells bind to 4773 unique host coding RNAs. Nine SARS-CoV-2 proteins upregulate target gene expression, including NSP12 and ORF9c, whose RNA substrates are associated with pathways in protein N-linked glycosylation ER processing and mitochondrial processes. Furthermore, siRNA knockdown of host genes targeted by viral proteins in human lung organoid cells identify potential antiviral host targets across different SARS-CoV-2 variants. Conversely, NSP9 inhibits host gene expression by blocking mRNA export and dampens cytokine productions, including interleukin-1alpha/beta. Our viral protein-RNA interactome provides a catalog of potential therapeutic targets and offers insight into the etiology of COVID-19 as a safeguard against future pandemics.