Abstract
The influenza virus RNA genome is transcribed and replicated in the context of the viral ribonucleoprotein (vRNP) complex by the viral RNA polymerase. The nucleoprotein (NP) is the structural component of the vRNP providing a scaffold for the viral RNA. In the vRNP as well as during transcription and replication the viral polymerase interacts with NP but it is unclear which parts of the polymerase and NP mediate these interactions. Previously the C-terminal ‘627’ domain (amino acids 538–693) of PB2 was shown to interact with NP. Here we report that a fragment encompassing amino acids 146–185 of NP is sufficient to mediate this interaction. Using NMR chemical shift perturbation assays we show that amino acid region 601 to 607 of the PB2 ‘627’ domain interacts with this fragment of NP. Substitutions of these PB2 amino acids resulted in diminished RNP activity and surface plasmon resonance assays showed that amino acids D605 was essential for the interaction with NP and V606 may also play a partial role in the interaction. Collectively these results reveal a possible interaction surface between NP and the PB2 subunit of the RNA polymerase complex.
Highlights
Influenza is a contagious respiratory disease that causes recurring annual epidemics and occasional pandemics
Each of the eight RNA segments of the virus is packaged into a viral ribonucleoprotein complex consisting of multiple nucleoproteins (NPs) and one heterotrimeric RNA polymerase, which binds the partially complementary 5’ and 3’ termini of the viral RNA (vRNA)
To identify putative amino acids in PB2 involved in the interaction with NP, we expressed 15N-lablled PB2 ‘627’ domain in bacterial expression system and performed a chemical shift perturbation (CSP) experiment, using either full-length NP or NP fragment 146–185 as ligand
Summary
Influenza is a contagious respiratory disease that causes recurring annual epidemics and occasional pandemics. According to the World Health Organization, influenza epidemics claim the life of 250,000 to 500,000 people every year. Influenza A virus genome comprises of eight segments of viral RNA (vRNA) encoding 10 major proteins and several auxiliary polypeptides [1,2]. Each of the eight RNA segments of the virus is packaged into a viral ribonucleoprotein (vRNP) complex consisting of multiple nucleoproteins (NPs) and one heterotrimeric RNA polymerase, which binds the partially complementary 5’ and 3’ termini of the vRNA.
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call