Fig. 2: Alternative polyadenylation of host transcripts is mediated by viral protein NS1 irrespective of amino-acids at positions 103/106 and abrogated by G184R mutation. | Nature Communications

Fig. 2: Alternative polyadenylation of host transcripts is mediated by viral protein NS1 irrespective of amino-acids at positions 103/106 and abrogated by G184R mutation.

From: mRNA 3’UTR lengthening by alternative polyadenylation attenuates inflammatory responses and correlates with virulence of Influenza A virus

Fig. 2

a Depicted viral proteins were transiently expressed in HEK293T cells via expression plasmid transfection (or transfection reagent only, vehicle), followed by quantification of APA of selected genes. ΔPDUI as a measure of APA status (left) and transgene transcript abundances (right) for 2 separately transfected wells are shown. b Schematic representation of previously described naming convention5 used in respect to NS1 mutants used in this study. c Transient expression of full length NS1 (NS1-wt, aa 1–230), NS1 effector domain (ED, aa 74–230), NS1 RNA-binding domain (RBD, aa 1–113) or transfection reagent only (vehicle) via expression plasmid transfection, followed by quantification of expression levels and APA of selected genes. ΔPDUI as a measure of APA status and NS1 transcript abundances are shown alongside mean for 2 separately transfected wells. d Amino acid sequences of IAV NS1 proteins (N = 35,326, NCBI Influenza virus database)), were aligned using Clustal Omega algorithm90. Normalized entropy (22-letter) as a measure of positional variation at indicated positions is depicted alongside sequence logos. e Indicated NS1 mutant proteins or controls (CTR – Thogotovirus M, vehicle – transfection reagent only) were transiently expressed in HEK293T cells via expression plasmid transfection, followed by quantification of transgene expression levels and APA of selected genes. ΔPDUI as a measure of APA status and transgene transcript abundances are shown alongside mean +/− sd for 3 separately transfected wells. Statistics refer to comparisons between indicated samples and gene-matched vehicle controls. f A549 cells were infected with indicated strains of IAV (based on PR8 background, MOI 3) for 24 h, followed by quantification of NS1 expression levels and APA of selected genes. ΔPDUI as a measure of APA status and NS1 transcript abundances are shown alongside mean +/− sd for 4 separately infected wells. Statistics refer to comparisons between indicated samples and gene-matched mock controls. Statistics were calculated using two-sided equal variance t-test. n.s. p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001. ΔCt values were calculated relative to the housekeeping gene RPLP0.

Back to article page