FIG. 2.
Association of Rpa34 and Rpa49 with Pol I in rpa34Δ or rpa49 mutants. ChIP assays were done on three independent cultures (comprising 100 ml of YPD) harvested at an A600 of 0.6. Pol I was revealed by polyclonal anti-Rpa190 and anti-Rpa34 or anti-Rpa49 rabbit antibodies (49). DNA occupancy was defined by the ratio between the immunoprecipitation (IP) signal and the DNA input (IN) signal. The diagram at the bottom shows the approximate positions of the primers used for reverse transcription-PCR (RT-PCR) amplification. Arrows denote the start sites and transcription orientations of the 5S and 35S rRNA transcripts. NTS1 and NTS2, nontranscribed spacers 1 and 2; nt, nucleotides. (Left panels) Effect of rpa49 mutations on the binding of Rpa34 to Pol I. Strain D704-6C was transformed with pVV200 plasmids bearing the mutant (rpa49-119,416) and wild-type (WT) RPA49 alleles. (Right panels) Effect of rpa34Δ on the binding of Rpa49 in YPH499 (WT) and T4-1C (rpa34Δ).