Methylation of RNA Cap in SARS-CoV-2 captured by serial crystallography

The genome of the SARS-CoV-2 coronavirus contains 29 proteins, of which 15 are nonstructural. Nsp10 and Nsp16 form a complex responsible for the capping of mRNA at the 5′ terminus. In the methylation reaction the S-adenosyl-L-methionine serves as the donor of the methyl group that is transferred to...

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Bibliographic Details
Published in:bioRxiv 2020-08
Main Authors: Wilamowski, Mateusz, Sherrell, Darren, Minasov, George, Kim, Youngchang, Shuvalova, Ludmilla, Lavens, Alex, Chard, Ryan, Maltseva, Natalia, Jedrzejczak, Robert, Rosas-Lemus, Monica, Saint, Nicklaus, Foster, Ian T, Michalska, Karolina, Satchell, Karla J, Joachimiak, Andrzej
Format: Article
Language:English
Subjects:
Coronaviruses
Crystallography
Crystals
Data collection
DNA methylation
Genomes
Methionine
Methyltransferase
Molecular Biology
mRNA guanylyltransferase
Ribose
Severe acute respiratory syndrome coronavirus 2
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Summary:The genome of the SARS-CoV-2 coronavirus contains 29 proteins, of which 15 are nonstructural. Nsp10 and Nsp16 form a complex responsible for the capping of mRNA at the 5′ terminus. In the methylation reaction the S-adenosyl-L-methionine serves as the donor of the methyl group that is transferred to Cap-0 at the first transcribed nucleotide to create Cap-1. The presence of Cap-1 makes viral RNAs mimic the host transcripts and prevents their degradation. To investigate the 2′- O methyltransferase activity of SARS-CoV-2 Nsp10/16, we applied fixed-target serial synchrotron crystallography (SSX) which allows for physiological temperature data collection from thousands of crystals, significantly reducing the x-ray dose while maintaining a biologically relevant temperature. We determined crystal structures of Nsp10/16 that revealed the states before and after the methylation reaction, for the first time illustrating coronavirus Nsp10/16 complexes with the m7GpppAm2′-O Cap-1, where 2′OH of ribose is methylated. We compare these structures with structures of Nsp10/16 at 297 K and 100 K collected from a single crystal. This data provide important mechanistic insight and can be used to design small molecules that inhibit viral RNA maturation making SARS-CoV-2 sensitive to host innate response. Competing Interest Statement The authors have declared no competing interest. Footnotes * Two authors had typos in their names, they were corrected.
ISSN:2692-8205
DOI:10.1101/2020.08.14.251421