PubMed Journals: Nucleic Acids Res
Source: PMID: 31131400
⇦ ⇨ Nucleic Acids Res. 2019 Jul 9;47(12):6538-6550.
⇩ doi: 10.1093/nar/gkz409.
Delicate structural coordination of the
Severe Acute Respiratory Syndrome coronavirus
Nsp13 upon ATP hydrolysis.
Jia Z(1), Yan L(1), Ren Z(2), Wu L(3), Wang
J(4), Guo J(5), Zheng L(1), Ming Z(6), Zhang
L(1), Lou Z(1), Rao Z(1)(2)(3).
(1) Laboratory of Structural Biology, School
of Medicine, Tsinghua University, Beijing
(2) State Key Laboratory of Medicinal Chemical
Biology, College of Life Science, Nankai
University, Tianjin 300353, China.
(3) Shanghai Institute for Advanced Immunochemical
Studies and iHuman Institute, ShanghaiTech
University, Shanghai 201210, China.
(4) State Key Laboratory of Biotherapy,
West China Hospital, Sichuan University,
Chengdu 610041, China.
(5) Protein Chemistry Facility, Center for
Biomedical Analysis of Tsinghua University,
Beijing 100084, China.
(6) State Key Laboratory of Conservation
and Utilization of Subtropical Agro-Bioresources,
College of Life Science and Technology,
Guangxi University, Nanning, China.
To date, an effective therapeutic treatment
that confers strong attenuation toward coronaviruses
(CoVs) remains elusive. Of all the potential
drug targets, the helicase of CoVs is considered
to be one of the most important. Here, we
first present the structure of the full-length
Nsp13 helicase of SARS-CoV (SARS-Nsp13) and
investigate the structural coordination
of its five domains and how these contribute
to its translocation and unwinding activity.
A translocation model is proposed for the
Upf1-like helicase members according to
three different structural conditions in
solution characterized through H/D exchange
assay, including substrate state (SARS-Nsp13-dsDNA
bound with AMPPNP), transition state (bound with ADP-AlF4-)
and product state (bound with ADP). We observed
that the β19-β20 loop on the 1A domain is
involved in unwinding process directly.
Furthermore, we have shown that the RNA
dependent RNA polymerase (RdRp), SARS-Nsp12,
can enhance the helicase activity of SARS-Nsp13
through interacting with it directly. The interacting
regions were identified and can be considered
common across CoVs, which provides new insights
into the Replication and Transcription Complex
(RTC) of CoVs.
© The Author(s) 2019. Published by Oxford
University Press on behalf of Nucleic Acids
DOI: 10.1093/nar/gkz409 PMCID: PMC6614802