tetano
Editor, Senior Moderator
Int J Biochem Cell Biol. 2014 May 27. pii: S1357-2725(14)00180-0. doi: 10.1016/j.biocel.2014.05.022. [Epub ahead of print]
Biophysical characterization of sites of host adaptive mutation in the influenza A virus RNA polymerase PB2 RNA-binding domain.
Lim K1, Kim M2, Lee MK1, Ko J1, Hong S1, Choi BS3.
Author information
Abstract
Influenza RNA polymerase is composed of three subunits, PA, PB1, and PB2, which interact with each other for transcription and replication of the viral RNA genome in the nucleus of infected cells. PB2 RNA-binding 627-domain (residues 535-693), located in the C-terminus, presents a highly basic surface around residue lysine 627 and has been proposed to interact with viral or cellular factors, resulting in host adaptation. However, the function of this domain is not yet characterized in detail. In this study, we identified RNA-binding activity and RNA-binding surfaces in both the N-terminal and basic C-terminal regions of PB2 627-domain using NMR experiments. Through mutagenesis studies, we confirmed which residues directly interact with RNA and mapped their locations on the RNA-binding surface. In addition, by luciferase activity assays, we showed that influenza virus polymerase activity may correlate with the interaction between PB2 and RNA. Representative host adaptive mutations (residues 591 and 627) were found to be located on the RNA-binding surface and were confirmed to directly interact with RNA and to affect polymerase activity. From these results, we suggest that influenza virus polymerase activity may be regulated through the interaction between PB2 627-domain and RNA and that consequently host adaptation of the virus may be influenced.
Copyright ? 2014. Published by Elsevier Ltd.
KEYWORDS:
Host adaptive mutation; Influenza RNA polymerase; Nuclear magnetic resonance; PB2 subunit; RNA-binding activity
PMID:
24875650
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24875650
Biophysical characterization of sites of host adaptive mutation in the influenza A virus RNA polymerase PB2 RNA-binding domain.
Lim K1, Kim M2, Lee MK1, Ko J1, Hong S1, Choi BS3.
Author information
Abstract
Influenza RNA polymerase is composed of three subunits, PA, PB1, and PB2, which interact with each other for transcription and replication of the viral RNA genome in the nucleus of infected cells. PB2 RNA-binding 627-domain (residues 535-693), located in the C-terminus, presents a highly basic surface around residue lysine 627 and has been proposed to interact with viral or cellular factors, resulting in host adaptation. However, the function of this domain is not yet characterized in detail. In this study, we identified RNA-binding activity and RNA-binding surfaces in both the N-terminal and basic C-terminal regions of PB2 627-domain using NMR experiments. Through mutagenesis studies, we confirmed which residues directly interact with RNA and mapped their locations on the RNA-binding surface. In addition, by luciferase activity assays, we showed that influenza virus polymerase activity may correlate with the interaction between PB2 and RNA. Representative host adaptive mutations (residues 591 and 627) were found to be located on the RNA-binding surface and were confirmed to directly interact with RNA and to affect polymerase activity. From these results, we suggest that influenza virus polymerase activity may be regulated through the interaction between PB2 627-domain and RNA and that consequently host adaptation of the virus may be influenced.
Copyright ? 2014. Published by Elsevier Ltd.
KEYWORDS:
Host adaptive mutation; Influenza RNA polymerase; Nuclear magnetic resonance; PB2 subunit; RNA-binding activity
PMID:
24875650
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24875650