The RNA binding domain of the Hantaan virus N protein maps to a central, conserved region

Xiaolin Xu, William Severson, Noah Villegas, Connie S. Schmaljohn, Colleen Jonsson

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

The nucleocapsid (N) protein of hantaviruses encapsidates both viral genomic and antigenomic RNAs, although only the genomic viral RNA (vRNA) is packaged into virions. To define the domain within the Hantaan virus (HTNV) N protein that mediates these interactions, 14 N- and C-terminal deletion constructs were cloned into a bacterial expression vector, expressed, and purified to homogeneity. Each protein was examined for its ability to bind the HTNV S segment vRNA with filter binding and gel electrophoretic mobility shift assays. These studies mapped a minimal region within the HTNV N protein (amino acids 175 to 217) that bound vRNA. Sequence alignments made from several hantavirus N protein sequences showed that the region identified has a 58% identity and an 86% similarity among these amino acid sequences. Two peptides corresponding to amino acids 175 to 196 (N1) and 197 to 218 (N2) were synthesized. The RNA binding of each peptide was measured by filter binding and competition analysis. Three oligoribonucleotides were used to measure binding affinity and assess specificity. The N2 peptide contained the major RNA binding determinants, while the N1 peptide, when mixed with N2, contributed to the specificity of vRNA recognition.

Original languageEnglish (US)
Pages (from-to)3301-3308
Number of pages8
JournalJournal of Virology
Volume76
Issue number7
DOIs
StatePublished - Mar 25 2002

Fingerprint

Hantaan virus
Viral RNA
RNA
Hantavirus
Peptides
peptides
proteins
Oligoribonucleotides
Nucleocapsid Proteins
Amino Acids
Sequence Alignment
Electrophoretic Mobility Shift Assay
oligoribonucleotides
Virion
amino acid sequences
Amino Acid Sequence
Proteins
Gels
genomics
nucleocapsid

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

The RNA binding domain of the Hantaan virus N protein maps to a central, conserved region. / Xu, Xiaolin; Severson, William; Villegas, Noah; Schmaljohn, Connie S.; Jonsson, Colleen.

In: Journal of Virology, Vol. 76, No. 7, 25.03.2002, p. 3301-3308.

Research output: Contribution to journalArticle

Xu, Xiaolin ; Severson, William ; Villegas, Noah ; Schmaljohn, Connie S. ; Jonsson, Colleen. / The RNA binding domain of the Hantaan virus N protein maps to a central, conserved region. In: Journal of Virology. 2002 ; Vol. 76, No. 7. pp. 3301-3308.
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