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Changing S158 to glutamic acid reduces the binding of HEXIM1 and P-TEFb to 7SK snRNA in vitro. Recombinant GST-Hex, GST-HexS158A and GST-HexS158E proteins alone or with P-TEFb were incubated with α-32P-labeled 7SK snRNA. RNA–protein complexes were separated by non-denaturing PAGE and visualized by autoradiography. Arrows to the left indicate the free 7SK snRNA, HEXIM1-7SK snRNA complex or 7SK snRNP.
