tetano
Editor, Senior Moderator
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[SUP] 1 2 3 [/SUP], Darren A Sherrell[SUP] 4 [/SUP], George Minasov[SUP] 5 [/SUP], Youngchang Kim[SUP] 1 4 [/SUP], Ludmilla Shuvalova[SUP] 5 [/SUP], Alex Lavens[SUP] 4 [/SUP], Ryan Chard[SUP] 6 [/SUP], Natalia Maltseva[SUP] 1 4 [/SUP], Robert Jedrzejczak[SUP] 1 4 [/SUP], Monica Rosas-Lemus[SUP] 5 [/SUP], Nickolaus Saint[SUP] 6 [/SUP], Ian T Foster[SUP] 6 [/SUP], Karolina Michalska[SUP] 1 4 [/SUP], Karla J F Satchell[SUP] 5 [/SUP], Andrzej Joachimiak[SUP] 7 2 4 [/SUP]
Affiliations
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 ([SUP]m7[/SUP]GpppA[SUB]m2'-O[/SUB]). 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.
. 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[SUP] 1 2 3 [/SUP], Darren A Sherrell[SUP] 4 [/SUP], George Minasov[SUP] 5 [/SUP], Youngchang Kim[SUP] 1 4 [/SUP], Ludmilla Shuvalova[SUP] 5 [/SUP], Alex Lavens[SUP] 4 [/SUP], Ryan Chard[SUP] 6 [/SUP], Natalia Maltseva[SUP] 1 4 [/SUP], Robert Jedrzejczak[SUP] 1 4 [/SUP], Monica Rosas-Lemus[SUP] 5 [/SUP], Nickolaus Saint[SUP] 6 [/SUP], Ian T Foster[SUP] 6 [/SUP], Karolina Michalska[SUP] 1 4 [/SUP], Karla J F Satchell[SUP] 5 [/SUP], Andrzej Joachimiak[SUP] 7 2 4 [/SUP]
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 ([SUP]m7[/SUP]GpppA[SUB]m2'-O[/SUB]). 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.