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| Status |
Public on Dec 01, 2020 |
| Title |
The SARS-CoV-2 RNA-protein interactome in infected human cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Other
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| Summary |
Characterizing the interactions that SARS-CoV-2 viral RNAs make with host cell proteins during infection can improve our understanding of viral RNA functions and the host innate immune response. Using RNA antisense purification and mass spectrometry (RAP-MS), we identify up to 104 human proteins that directly and specifically bind to SARS-CoV-2 RNAs in infected human cells. We integrate the SARS-CoV-2 RNA interactome with proteome abundance changes induced by viral infection and link interactome proteins to cellular pathways relevant to SARS-CoV-2 infections. We demonstrate by genetic perturbation that CNBP and LARP1, two of the most strongly enriched viral RNA binders, restrict SARS-CoV-2 replication in infected cells and provide a global map of their direct RNA contact sites. Pharmacological inhibition of three other RNA interactome members, PPIA, ATP1A1, and the ARP2/3 complex, reduces viral replication in two human cell lines. The identification of host dependency factors and defence strategies as presented in this work will improve the design of targeted therapeutics against SARS-CoV-2.
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| Overall design |
1. We mapped direct protein-RNA interaction sites using enhanced crosslinking and immunoprecipitation (eCLIP). 2. We identified RNA sequences crosslinked to proteins purified by RNA antisense purification (RAP-MS) by following the procedure described in the protocols section.
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| Contributor(s) |
Munschauer M, Lareau C |
| Citation(s) |
33349665 |
| Submission date |
Jul 14, 2020 |
| Last update date |
Mar 02, 2021 |
| Contact name |
Caleb Lareau |
| E-mail(s) |
lareauc@mskcc.org
|
| Organization name |
Memorial Sloan Kettering
|
| Street address |
417 E 68th St, Zuckerman - ZRC 1132
|
| City |
New York |
| State/province |
New York |
| ZIP/Postal code |
10065 |
| Country |
USA |
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| Platforms (2) |
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| Samples (10)
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| Relations |
| BioProject |
PRJNA646247 |
| SRA |
SRP271809 |
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