Figure 7.

hnRNP A2 binds the first 71 nt of human telomerase RNA. (A) ³²P-labeled hTR 5′ 71-nt was incubated with increasing amounts (0, 0.1, 0.5, 1, 2, 5 and 10 pmol) of recombinant hnRNP A2, and separated on a 15-cm 5% polyacrylamide gel. These assays were performed in the presence of 1 μg/μl heparin. The arrow on the right marks the retarded RNA–protein complex. (B) Assay was performed as above, except that the RNA used was first denatured by heating to 95°C for 5 min, cooled by placing immediately on ice and then incubated with protein. (C) ³²P-labeled hTR 5′ 71-nt RNA was incubated with 1, 10 and 100 μM of each recombinant protein, and separated on a 15-cm 5% native polyacrylamide gel. The single and double arrowheads point to the monomer and dimer forms of the RNA, respectively.