FIG. 1.

p65 increases the amount of RNA associated with TERT and the enzymatic activity of telomerase. (A) A cartoon of Tetrahymena TER with secondary structural elements indicated. TBE, template boundary element or TERT-binding element; TRE, template recognition element. (B) An SDS polyacrylamide gel stained with Coomassie blue showing the purity of p65. (C) Direct activity assay performed on wild-type telomerase synthesized in the absence or presence of enough purified p65 to saturate TER. RC, 5′-³²P-end-labeled DNA oligonucleotide added as a recovery control. Telomerase repeats added are indicated on the right side of the gel. Rel act, radiolabeled nucleotide incorporation relative to that in the absence of p65. (D) Duplicate SDS-PAGE analysis of TERT immunopurification of telomerase assembled in RRLs in the presence or absence of p65 (left) and the TERT-independent recovery of TER (right). The top panel shows the [³⁵S]methionine incorporated into TERT and the coimmunoprecipitated ³²P-labeled TER. The bottom panel is an exposure of the same gel with film inserted to block the detection of ³⁵S, showing only the coimmunoprecipitation of ³²P-labeled TER. Rel RNA, relative amount of TER pulldown normalized to the amount of TERT.