Fig. 4: A nuclear incompatibility determinant impairs phage fitness.
From: Viral speciation through subcellular genetic isolation and virogenesis incompatibility
a Efficiency of plating (EOP) relative to GFPmut1 for ΦPA3 in cells expressing the indicated fusions with 1% arabinose. The number of independent replicates, n, equals 4 (GFPmut1), 3 (sfGFP), 4 (gp210-GFPmut1), and 4 (gp210-sfGFP). Error bars equal standard deviation. b EOP relative to GFPmut1 for ΦKZ in cells expressing the indicated fusions with 1% arabinose demonstrate a 99.4% decrease in viable ΦKZ with gp210-GFPmut1 in the nucleus. The number of independent replicates, n, equals 7 (GFPmut1), 6 (sfGFP), 6 (gp210-GFPmut1), 6 (gp210-sfGFP). Error bars equal standard deviation. c, d GFP fusions (green) and FM4-64 stained cell membranes (red). Individual data points are shown as circles. c GFPmut1 and sfGFP localize to the cytoplasm during ΦPA3 infections. gp210-GFPmut1 and gp210-sfGFP localize to the nucleus and form puncta during ΦPA3 infections. d sfGFP and gp210-sfGFP localize to the cytoplasm of ΦKZ infected cells. GFPmut1 and gp210-GFPmut1 localize to the nucleus of ΦKZ infected cells. e–h Determination of ΦKZ IC50 (red dotted line) for cells expressing control proteins GFPmut1 or sfGFP (e, f) or fusions to gp210 (g, h) as indicated by measuring cell growth (OD600) over 6.5 h of infection. Ten-fold serial dilutions of phage were added to cells resulting in a multiplicity of infection (MOI) ranging from 5000 to 5 × 10−7 as shown in (d). All growth curves represent an average of eight independent trials. For the sake of clarity, only the MOIs that indicate the IC50 are shown for (f–h). Source data are provided as a Source Data file.
