Figure 1.
The hDbr1 protein associates with xeroderma pigmentosum, complementeation group A (XPA)-binding protein 2 (Xab2) in vivo and in vitro. (A) The in vivo association of several IL complex proteins with hDbr1 protein was analyzed by immunoprecipitation assay. HEK293T whole-cell extract was prepared after transfection with Flag-vector, Flag-hPrp19, Flag-Xab2, Flag-hCrn or Flag-IBP160 plasmid. Flag-tagged proteins were precipitated using anti-Flag M2 agarose from whole-cell extract, and the precipitates (IPed) were subjected to Western blotting analyses using anti-hDbr1 antibody (upper panel) and an anti-Flag polyclonal antibody (lower panel). A portion (5%) of the precipitated proteins (lanes marked as Input) was also immunoblotted. The position of endogenous hDbr1 protein is indicated by an arrow in the upper panel. The full-length Flag-tagged proteins are marked with asterisks in the lower panel. The positions of the protein size markers are described on the left of the panel; and (B) In vitro binding of hDbr1 to Xab2. Flag-tagged hPrp19, hCrn and Xab2 proteins, produced and labeled with [35S]methionine in vitro, were incubated with 5 μg each of GST or GST-hDbr1. After precipitation and washing, bound proteins were eluted, separated on 12.5% SDS-PAGE gel and visualized by fluorography. The lanes marked Input contain 10% of the total protein used in each binding reaction. The position of molecular mass markers is shown on the left of the panel.