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Examination of specific binding activity of packaging sequence RNAs to the SARS‐CoV nucleocapsid by using cell‐based in vivo assay for RNA‐Protein interaction
Author(s) -
Woo Jiwon,
Cho YongEun
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.lb223
Subject(s) - rna , biology , virology , rna binding protein , coronavirus , viral replication , computational biology , microbiology and biotechnology , virus , gene , genetics , covid-19 , medicine , infectious disease (medical specialty) , disease , pathology
Severe acute respiratory syndrome coronavirus (SARS‐CoV) is the etiological agent of SARS, an emerging disease characterized by atypical pneumonia. The SARS‐CoV nucleocapsid protein serves multiple functions in viral replication, transcription and the formation of viral genome complex. Coronaviruses specifically package genomic RNA into assembled virions, little is known about the specific interaction between Nucleocapsid protein and packaging RNA sequence in previously. Herein, we exploited a cell‐based translation inhibition assay to elucidate the protein‐RNA interaction in living cells. Our results demonstrated that the packaging signal is pivotal in the specific interaction of the nucleocapsid and viral genomic RNA for incorporation into virions. We identified that the specific conserved RNA sequence of the nsp12 encoding vRNA of SARS‐CoV, and conserved RNAs of other corona viruses are major determinant sequences for the N‐protein interaction, respectively. In addition, using deletion mapping of the N‐protein, we also found that the specific in vivo RNA binding activity of the Nucleocapsid protein of the SARS‐CoV requires both the N‐terminal and C‐terminal domain. These results provide the first evidence for a specific interaction of the SARS‐CoV Nucleocapsid protein with the its viral packaging RNA sequence in vivo that will be useful as a valuable research tool for further detailed studies and providing new targets for antiviral therapies. Support or Funding Information This work was supported by the Brain Korea 21 PLUS Project for Medical Science, Yonsei University. In addition, this work was supported by a grant from the National Research Foundation of Korea (NRF‐2017R1D1A1B03030315). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .