Proc Natl Acad Sci U S A . 2'-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography

[tetano]

Proc Natl Acad Sci U S A


. 2021 May 25;118(21):e2100170118.
doi: 10.1073/pnas.2100170118.
2'-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography


Mateusz Wilamowski ^{1 2 3} , Darren A Sherrell ⁴ , George Minasov ⁵ , Youngchang Kim ^{1 4} , Ludmilla Shuvalova ⁵ , Alex Lavens ⁴ , Ryan Chard ⁶ , Natalia Maltseva ^{1 4} , Robert Jedrzejczak ^{1 4} , Monica Rosas-Lemus ⁵ , Nickolaus Saint ⁶ , Ian T Foster ⁶ , Karolina Michalska ^{1 4} , Karla J F Satchell ⁵ , Andrzej Joachimiak ^{7 2 4}



Affiliations

• PMID: 33972410
• DOI: 10.1073/pnas.2100170118

Abstract

The genome of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) coronavirus has a capping modification at the 5'-untranslated region (UTR) to prevent its degradation by host nucleases. These modifications are performed by the Nsp10/14 and Nsp10/16 heterodimers using S-adenosylmethionine as the methyl donor. Nsp10/16 heterodimer is responsible for the methylation at the ribose 2'-O position of the first nucleotide. To investigate the conformational changes of the complex during 2'-O methyltransferase activity, we used a fixed-target serial synchrotron crystallography method at room temperature. We determined crystal structures of Nsp10/16 with substrates and products that revealed the states before and after methylation, occurring within the crystals during the experiments. Here we report the crystal structure of Nsp10/16 in complex with Cap-1 analog (^m7GpppA_m2'-O). Inhibition of Nsp16 activity may reduce viral proliferation, making this protein an attractive drug target.

Keywords: CAP-1; Nsp10/16; SARS-CoV-2; mRNA; serial crystallography.