2′-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography

Mateusz Wilamowski; Darren A. Sherrell; George Minasov; Youngchang Kim; Ludmilla Shuvalova; Alex Lavens; Ryan Chard; Natalia Maltseva; Robert Jedrzejczak; Monica Rosas-Lemus; Nickolaus Saint; Ian T. Foster; Karolina Michalska; Karla J.F. Satchell; Andrzej Joachimiak

doi:10.1073/pnas.2100170118

2′-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography

Mateusz Wilamowski, Darren A. Sherrell, George Minasov, Youngchang Kim, Ludmilla Shuvalova, Alex Lavens, Ryan Chard, Natalia Maltseva, Robert Jedrzejczak, Monica Rosas-Lemus, Nickolaus Saint, Ian T. Foster, Karolina Michalska, Karla J.F. Satchell, Andrzej Joachimiak^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

50 Scopus citations

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.

Original language	English (US)
Article number	e2100170118
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	118
Issue number	21
DOIs	https://doi.org/10.1073/pnas.2100170118
State	Published - May 25 2021

Funding

ACKNOWLEDGMENTS. Funding for this research was provided in part by federal funds from the National Institute of Allergy and Infectious Diseases, NIH, Department of Health and Human Services, under Contract HHSN272201700060C and by the Department of Energy (DOE) Office of Science through the National Virtual Biotechnology Laboratory, a consortium of DOE national laboratories focused on response to COVID-19, with funding provided by the CARES Act. The use of SBC beamlines at the Advanced Photon Source and resources of the Argonne Leadership Computing Facility is supported by the DOE Office of Science and operated for the DOE Office of Science by the Argonne National Laboratory under Contract DE-AC02-06CH11357.

Keywords

Nsp10/16 | SARS-CoV-2 | mRNA | CAP-1 | serial crystallography

ASJC Scopus subject areas

General

Access to Document

10.1073/pnas.2100170118

Cite this

Wilamowski, M., Sherrell, D. A., Minasov, G., Kim, Y., Shuvalova, L., Lavens, A., Chard, R., Maltseva, N., Jedrzejczak, R., Rosas-Lemus, M., Saint, N., Foster, I. T., Michalska, K., Satchell, K. J. F., & Joachimiak, A. (2021). 2′-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography. Proceedings of the National Academy of Sciences of the United States of America, 118(21), Article e2100170118. https://doi.org/10.1073/pnas.2100170118

@article{d517ed5932184e5e92e46a707cf9318a,

title = "2′-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography",

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 (m7GpppAm2′-O). Inhibition of Nsp16 activity may reduce viral proliferation, making this protein an attractive drug target.",

keywords = "Nsp10/16 | SARS-CoV-2 | mRNA | CAP-1 | serial crystallography",

author = "Mateusz Wilamowski and Sherrell, {Darren A.} and George Minasov and Youngchang Kim and Ludmilla Shuvalova and Alex Lavens and Ryan Chard and Natalia Maltseva and Robert Jedrzejczak and Monica Rosas-Lemus and Nickolaus Saint and Foster, {Ian T.} and Karolina Michalska and Satchell, {Karla J.F.} and Andrzej Joachimiak",

year = "2021",

month = may,

day = "25",

doi = "10.1073/pnas.2100170118",

language = "English (US)",

volume = "118",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "National Academy of Sciences",

number = "21",

}

Wilamowski, M, Sherrell, DA, Minasov, G, Kim, Y, Shuvalova, L, Lavens, A, Chard, R, Maltseva, N, Jedrzejczak, R, Rosas-Lemus, M, Saint, N, Foster, IT, Michalska, K, Satchell, KJF & Joachimiak, A 2021, '2′-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography', Proceedings of the National Academy of Sciences of the United States of America, vol. 118, no. 21, e2100170118. https://doi.org/10.1073/pnas.2100170118

TY - JOUR

T1 - 2′-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography

AU - Wilamowski, Mateusz

AU - Sherrell, Darren A.

AU - Minasov, George

AU - Kim, Youngchang

AU - Shuvalova, Ludmilla

AU - Lavens, Alex

AU - Chard, Ryan

AU - Maltseva, Natalia

AU - Jedrzejczak, Robert

AU - Rosas-Lemus, Monica

AU - Saint, Nickolaus

AU - Foster, Ian T.

AU - Michalska, Karolina

AU - Satchell, Karla J.F.

AU - Joachimiak, Andrzej

PY - 2021/5/25

Y1 - 2021/5/25

N2 - 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 (m7GpppAm2′-O). Inhibition of Nsp16 activity may reduce viral proliferation, making this protein an attractive drug target.

AB - 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 (m7GpppAm2′-O). Inhibition of Nsp16 activity may reduce viral proliferation, making this protein an attractive drug target.

KW - Nsp10/16 | SARS-CoV-2 | mRNA | CAP-1 | serial crystallography

UR - http://www.scopus.com/inward/record.url?scp=85105756625&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85105756625&partnerID=8YFLogxK

U2 - 10.1073/pnas.2100170118

DO - 10.1073/pnas.2100170118

M3 - Article

C2 - 33972410

AN - SCOPUS:85105756625

SN - 0027-8424

VL - 118

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 21

M1 - e2100170118

ER -

2′-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography

Abstract

Funding

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Room Temperature Crystal Structure of Nsp10/Nsp16 from SARS-CoV-2 with Substrates and Products of 2'-O-methylation of the Cap-1

Room Temperature Structure of SARS-CoV-2 Nsp10/Nsp16 Methyltransferase in a Complex with 2'-O-methylated m7GpppA Cap-1 and SAH Determined by Fixed-Target Serial Crystallography

Room Temperature Structure of SARS-CoV-2 NSP10/NSP16 Methyltransferase in a Complex with SAM Determined by Fixed-Target Serial Crystallography

Cite this

2′-O methylation of RNA cap in SARS-CoV-2 captured by serial crystallography

Abstract

Funding

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Datasets

Room Temperature Crystal Structure of Nsp10/Nsp16 from SARS-CoV-2 with Substrates and Products of 2'-O-methylation of the Cap-1

Room Temperature Structure of SARS-CoV-2 Nsp10/Nsp16 Methyltransferase in a Complex with 2'-O-methylated m7GpppA Cap-1 and SAH Determined by Fixed-Target Serial Crystallography

Room Temperature Structure of SARS-CoV-2 NSP10/NSP16 Methyltransferase in a Complex with SAM Determined by Fixed-Target Serial Crystallography

Cite this