tetano
Editor, Senior Moderator
J Virol. 2012 Apr 24. [Epub ahead of print]
Sequence in the Influenza A Virus Nucleoprotein Required for Viral Polymerase Binding and RNA Synthesis.
Marklund JK, Ye Q, Dong J, Tao YJ, Krug RM.
Source
Department of Molecular Genetics and Microbiology, Institute for Cell and Molecular Biology, University of Texas at Austin, Texas USA.
Abstract
Many proposed mechanisms for influenza A viral RNA synthesis include an interaction of the nucleoprotein (NP) with the viral polymerase. To identify a NP sequence required for this interaction, we used the cryo-electron microscopic structure of a influenza virus mini-ribonucleoprotein as a guide for choosing promising surface-exposed sequences. We show that three amino acids (R204, W207, R208) located in a loop at the top of the head domain of NP are required for functional interaction with the viral polymerase. Quantitative RT-PCR measurements of RNAs synthesized in minigenome assays established that each of these NP amino acids is required for viral RNA synthesis. Mutation of these three amino acids does not affect nuclear localization, or RNA-binding and oligomerization activities of NP. In vitro binding experiments with purified virus polymerase and NPs established that these three amino acids are required for NP binding to the viral polymerase.
PMID:
22532672
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22532672
Sequence in the Influenza A Virus Nucleoprotein Required for Viral Polymerase Binding and RNA Synthesis.
Marklund JK, Ye Q, Dong J, Tao YJ, Krug RM.
Source
Department of Molecular Genetics and Microbiology, Institute for Cell and Molecular Biology, University of Texas at Austin, Texas USA.
Abstract
Many proposed mechanisms for influenza A viral RNA synthesis include an interaction of the nucleoprotein (NP) with the viral polymerase. To identify a NP sequence required for this interaction, we used the cryo-electron microscopic structure of a influenza virus mini-ribonucleoprotein as a guide for choosing promising surface-exposed sequences. We show that three amino acids (R204, W207, R208) located in a loop at the top of the head domain of NP are required for functional interaction with the viral polymerase. Quantitative RT-PCR measurements of RNAs synthesized in minigenome assays established that each of these NP amino acids is required for viral RNA synthesis. Mutation of these three amino acids does not affect nuclear localization, or RNA-binding and oligomerization activities of NP. In vitro binding experiments with purified virus polymerase and NPs established that these three amino acids are required for NP binding to the viral polymerase.
PMID:
22532672
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22532672