Essential amino acids of the Hantaan virus N protein in its interaction with RNA

William Severson, Xiaolin Xu, Michaela Kuhn, Nina Senutovitch, Mercy Thokala, François Ferron, Sonia Longhi, Bruno Canard, Colleen Jonsson

Research output: Contribution to journalArticle

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Abstract

The nucleocapsid (N) protein of hantavirus encapsidates viral genomic and antigenomic RNAs. Previously, deletion mapping identified a central, conserved region (amino acids 175 to 217) within the Hantaan virus (HTNV) N protein that interacts with a high affinity with these viral RNAs (vRNAs). To further define the boundaries of the RNA binding domain (RBD), several peptides were synthesized and examined for the ability to bind full-length S-segment vRNA. Peptide 195-217 retained 94% of the vRNA bound by the HTNV N protein, while peptides 175-186 and 205-217 bound only 1% of the vRNA. To further explore which residues were essential for binding vRNA, we performed a comprehensive mutational analysis of the amino acids in the RBD. Single and double Ala substitutions were constructed for 18 amino acids from amino acids 175 to 217 in the full-length N protein. In addition, Ala substitutions were made for the three R residues in peptide 185-217. An analysis of protein-RNA interactions by electrophoretic mobility shift assays implicated E192, Y206, and S217 as important for binding. Chemical modification experiments showed that lysine residues, but not arginine or cysteine residues, contribute to RNA binding, which agreed with bioinformatic predictions. Overall, these data implicate lysine residues dispersed from amino acids 175 to 429 of the protein and three amino acids located in the RBD as essential for RNA binding.

Original languageEnglish (US)
Pages (from-to)10032-10039
Number of pages8
JournalJournal of Virology
Volume79
Issue number15
DOIs
StatePublished - Aug 1 2005

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Hantaan virus
Essential Amino Acids
essential amino acids
Viral RNA
RNA
Amino Acids
Peptides
proteins
Lysine
amino acids
peptides
Hantavirus
Nucleocapsid Proteins
Proteins
Electrophoretic Mobility Shift Assay
Computational Biology
Cysteine
Arginine
Hantaan virus nucleocapsid protein
lysine

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Essential amino acids of the Hantaan virus N protein in its interaction with RNA. / Severson, William; Xu, Xiaolin; Kuhn, Michaela; Senutovitch, Nina; Thokala, Mercy; Ferron, François; Longhi, Sonia; Canard, Bruno; Jonsson, Colleen.

In: Journal of Virology, Vol. 79, No. 15, 01.08.2005, p. 10032-10039.

Research output: Contribution to journalArticle

Severson, W, Xu, X, Kuhn, M, Senutovitch, N, Thokala, M, Ferron, F, Longhi, S, Canard, B & Jonsson, C 2005, 'Essential amino acids of the Hantaan virus N protein in its interaction with RNA', Journal of Virology, vol. 79, no. 15, pp. 10032-10039. https://doi.org/10.1128/JVI.79.15.10032-10039.2005
Severson W, Xu X, Kuhn M, Senutovitch N, Thokala M, Ferron F et al. Essential amino acids of the Hantaan virus N protein in its interaction with RNA. Journal of Virology. 2005 Aug 1;79(15):10032-10039. https://doi.org/10.1128/JVI.79.15.10032-10039.2005
Severson, William ; Xu, Xiaolin ; Kuhn, Michaela ; Senutovitch, Nina ; Thokala, Mercy ; Ferron, François ; Longhi, Sonia ; Canard, Bruno ; Jonsson, Colleen. / Essential amino acids of the Hantaan virus N protein in its interaction with RNA. In: Journal of Virology. 2005 ; Vol. 79, No. 15. pp. 10032-10039.
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