The binding of 3AB1 at the bottom of the palm subdomain of 3Dpol increases localization of polymerase to intracellular membranes [Dataset]

Abstract

(A) Scheme representing the fluorescence microscopy experiments comparing the distribution of 3Dpol in the presence of 3AB1, wild type and mutant. The left panel shows 3Dpol interacting with the 3AB1 precursor bound to the membrane. The right panel mimics the scenario in the presence of the 3AB1 mutant 3AB1(P6S/R9A/R16A/L19S), unable to bind 3Dpol. (B) Fluorescence images of HeLa cells showing the different distribution of 3Dpol bound to 3AB1 wild type or the 3AB1(P6A/R9A/R16A/L19S) mutant. Upper panels show the control cells transfected with the polymerase only (green), which appears distributed throughout the cell. Middle panels show cells transfected with 3AB1 wild type (red) and 3Dpol (green), where 3Dpol mostly co-localizes with the 3AB1 protein in a continuous compartment in the cytoplasm. The lower panels show cells transfected with the 3Dpol and the 3AB1 mutant (P6A/R9A/R16A/L19S), where 3Dpol recovers its localization throughout the cell. The images shown are representative of the total number of images obtained. (C) Fluorescence Intensity plots comparing the relative distribution of 3Dpol (green) in presence of wild type and mutant 3AB1 proteins (red), left and right panels, respectively. The nucleus is labelled in blue (DAPI).