FIG. 5.

Truncation mutants of telomere repeat factors inhibit OriP replication. (A) OriPwt or control plasmid ΔOriP was transfected into ZKO-293 cells with pCMV-Flag-2 vector or an expression plasmid for hRap1, TRF1, or TRF2 and their derivatives as indicated above each lane. Replicated DNA was measured by DpnI resistance in the top panel (DpnI/BamHI), and total extracted DNA is indicated in the bottom panel (BamHI digest). (B) PhosphorImager quantitation of at least three independent Southern blots is presented as a bar graph. (C) Transfected proteins were detected by Western blotting with anti-Flag antibody (full-length hRap1 is not Flag tagged), or the same blot was reprobed with anti-TRF2 (full-length TRF2 is not Flag tagged). (D) Schematic diagram of truncation mutants cloned into pCMV-Flag-2. The hRap1 BRCT (BRCA1 C-terminal homology), myb DNA binding, TRF2 interaction (TRF2-IX), and TRF homodimerization (TRFH) domains are indicated.