An in vivo cell-based assay for investigating the specific interaction between the SARS-CoV N-protein and its viral RNA packaging sequence.
Identifieur interne : 000B84 ( Main/Exploration ); précédent : 000B83; suivant : 000B85An in vivo cell-based assay for investigating the specific interaction between the SARS-CoV N-protein and its viral RNA packaging sequence.
Auteurs : Jiwon Woo [Corée du Sud] ; Eunice Yoojin Lee [États-Unis] ; Mirae Lee [Corée du Sud] ; Taeyeon Kim [Corée du Sud] ; Yong-Eun Cho [Corée du Sud]Source :
- Biochemical and biophysical research communications [ 1090-2104 ] ; 2019.
Abstract
The SARS-CoV nucleocapsid (N) protein serves multiple functions in viral replication, transcription, and assembly of the viral genome complex. Coronaviruses specifically package genomic RNA into assembled virions, and in SARS-CoV, it is reported that this process is driven by an interaction between the N-protein and a packaging signal encoded within the viral RNA. While recent studies have uncovered the sequence of this packaging signal, little is known about the specific interaction between the N-protein and the packaging signal sequence, and the mechanisms by which this interaction drives viral genome packaging. In this study, we developed a novel in vivo cell-based assay for examining this interaction between the N-protein and packaging signal RNA for SARS-CoV, as well as other viruses within the coronaviridae family. Our results demonstrate that the N-protein specifically recognizes the SARS-CoV packaging signal with greater affinity compared to signals from other coronaviruses or non-coronavirus species. We also use deletion mapping to identify a 151-nt region within the packaging signal sequence that is critical for N-protein-RNA binding, and conversely, we show that both the N-terminal and C-terminal domains of the N protein are necessary for recognizing the packaging RNA. These results describe, for the first time, in vivo evidence for an interaction between the SARS-CoV N-protein and its packaging signal RNA, and demonstrate the feasibility of using this cell-based assay to further probe viral RNA-protein interactions in future studies.
DOI: 10.1016/j.bbrc.2019.09.115
PubMed: 31594639
Affiliations:
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Electronic address: yecho@yuhs.ac.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, 03722, Republic of Korea; The Spine and Spinal Cord Institute, Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, 06273</wicri:regionArea><wicri:noRegion>06273</wicri:noRegion></affiliation></author></analytic><series><title level="j">Biochemical and biophysical research communications</title><idno type="eISSN">1090-2104</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The SARS-CoV nucleocapsid (N) protein serves multiple functions in viral replication, transcription, and assembly of the viral genome complex. Coronaviruses specifically package genomic RNA into assembled virions, and in SARS-CoV, it is reported that this process is driven by an interaction between the N-protein and a packaging signal encoded within the viral RNA. While recent studies have uncovered the sequence of this packaging signal, little is known about the specific interaction between the N-protein and the packaging signal sequence, and the mechanisms by which this interaction drives viral genome packaging. In this study, we developed a novel in vivo cell-based assay for examining this interaction between the N-protein and packaging signal RNA for SARS-CoV, as well as other viruses within the coronaviridae family. Our results demonstrate that the N-protein specifically recognizes the SARS-CoV packaging signal with greater affinity compared to signals from other coronaviruses or non-coronavirus species. We also use deletion mapping to identify a 151-nt region within the packaging signal sequence that is critical for N-protein-RNA binding, and conversely, we show that both the N-terminal and C-terminal domains of the N protein are necessary for recognizing the packaging RNA. These results describe, for the first time, in vivo evidence for an interaction between the SARS-CoV N-protein and its packaging signal RNA, and demonstrate the feasibility of using this cell-based assay to further probe viral RNA-protein interactions in future studies.</div></front></TEI><affiliations><list><country><li>Corée du Sud</li><li>États-Unis</li></country></list><tree><country name="Corée du Sud"><noRegion><name sortKey="Woo, Jiwon" sort="Woo, Jiwon" uniqKey="Woo J" first="Jiwon" last="Woo">Jiwon Woo</name></noRegion><name sortKey="Cho, Yong Eun" sort="Cho, Yong Eun" uniqKey="Cho Y" first="Yong-Eun" last="Cho">Yong-Eun Cho</name><name sortKey="Kim, Taeyeon" sort="Kim, Taeyeon" uniqKey="Kim T" first="Taeyeon" last="Kim">Taeyeon Kim</name><name sortKey="Lee, Mirae" sort="Lee, Mirae" uniqKey="Lee M" first="Mirae" last="Lee">Mirae Lee</name></country><country name="États-Unis"><noRegion><name sortKey="Lee, Eunice Yoojin" sort="Lee, Eunice Yoojin" uniqKey="Lee E" first="Eunice Yoojin" last="Lee">Eunice Yoojin Lee</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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