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. 2008 Aug 22;283(34):23016–23025. doi: 10.1074/jbc.M801021200

FIGURE 6.

FIGURE 6.

TbRGG2 is an RNA-binding protein. A, TbRGG2 binds to gRNA and displays a preference for poly(U). Five fmol of body-labeled gCYb[558] was incubated with 1 μm purified GST-TbRGG2 for 20 min (lane 2). Lane 1 is 1 μm GST-TbRGG2 in the absence of radiolabeled gRNA. For competitor analyses (lanes 3–6), 5 fmol (lane 3), 50 fmol (lane 4), 500 fmol (lane 5), or 5 pmol (lane 6) of the indicated cold homoribopolymer was added to 1 μm GST-TbRGG2 immediately prior to the addition to 5 fmol gCYb[558]. Reactions were subjected to UV cross-linking and treated with RNase A, and proteins were resolved by 10% SDS-PAGE. Proteins that were cross-linked to radiolabeled gCYb[558] was visualized by PhosphorImager analysis. B, TbRGG2 binds to A6 mRNA in vitro. Five fmol of body-labeled 3′A6U mRNA was incubated with 1 μm purified GST-TbRGG2 as in A. Lane 1, 3′A6U RNA in the absence of TbRGG2 protein. The indicated unlabeled competitor RNAs (lanes 3–5) were added as in A ina1× (5 fmol, lane 3), 10× (50 fmol, lane 4), or 100× molar excess (500 fmol, lane 5) immediately prior to the addition of 5 fmol of radiolabeled 3′A6U.