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Updated: Aug 15, 2026

Purification and Visualization of Influenza A Viral Ribonucleoprotein Complexes
Published on: February 9, 2009
Reconstitution of influenza virus RNA-nucleoprotein complexes structurally resembling native viral ribonucleoprotein
K Yamanaka1, A Ishihama, K Nagata
1Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka, Japan.
Abstract:
Reconstitution of influenza virus nucleoprotein (NP)-RNA complexes was performed with segment 8 RNA, which was synthesized in vitro from cDNA, and NP purified from virions. Under optimum conditions established using a filter binding assay and a gel retardation assay, NP was found to bind any RNA longer than 15 nucleotides. NP-RNA complexes formed at 30 degrees C are more resistant to high concentrations of NaCl than those formed at 0 degrees C. Treatment of NP with N-ethylmaleimide gave no effect on its RNA binding activity, whereas treatment with alkaline phosphatase enhanced its RNA binding activity. The newly developed "reverse-printing" method of RNase V1-treated complexes revealed that reconstituted NP-RNA complexes carry RNase V1-sensitive sites as do native ribonucleoprotein (RNP) cores (RNA polymerase-NP-RNA complexes), implying that RNA-NP complexes structurally similar to native RNP cores are reconstituted from isolated components.
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