Open AccessA statistical strategy to identify recombinant viral ribonucleoprotein of avian, human, and swine influenza A viruses with elevated polymerase activity
Author(s) -
Chin Alex W. H.,
Greenbaum Benjamin D.,
Li Olive T. W.,
Webby Richard J.,
Poon Leo L. M.
Publication year - 2013
Publication title -
influenza and other respiratory viruses
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.743
H-Index - 57
eISSN - 1750-2659
pISSN - 1750-2640
DOI - 10.1111/irv.12117
Subject(s) - ribonucleoprotein , reassortment , polymerase , biology , virology , rna dependent rna polymerase , influenza a virus , protein subunit , virus , microbiology and biotechnology , genetics , gene , rna , covid-19 , medicine , disease , pathology , infectious disease (medical specialty)
Objectives Reassortment of influenza A viruses can give rise to viral ribonucleoproteins (v RNP s) with elevated polymerase activity and the previous three pandemic influenza viruses contained reassorted v RNP s of different origins. These suggest that reassorted v RNP may be one of the factors leading to a pandemic virus. In this study, we reconstituted chimeric v RNP s with three different viral strains isolated from avian, human and swine hosts. We applied a statistical strategy to identify the effect that the origin of a single v RNP protein subunit or the interactions between these subunits on polymerase activity. Design Eighty one chimeric v RNP s were reconstituted in 293 T cells at different temperatures. Polymerase activity was determined by luciferase reporter assay and the results were analysed by multiway anova and other statistical methods. Results It was found that PB 2, PB 1, NP , PB 2‐ PB 1 interaction, PB 2‐ PA interaction and PB 1‐ NP interaction had significant effect on polymerase activity at 37°C and several single subunits and interactions were identified to lead to elevation of polymerase activity. Furthermore, we studied 27 out of these 81 different chimieric v RNP s in different combinations via fractional factorial design approach. Our results suggested that the approach can identify the major single subunit or interaction factors that affect the polymerase activity without the need to experimentally reproduce all possible v RNP combinations. Conclusions Statistical approach and fractional factorial design are useful to identify the major single subunit or interaction factors that can modulate viral polymerase activity.