Fragile DNA Motifs Trigger Mutagenesis at Distant Chromosomal Loci in Saccharomyces cerevisiae
Figure 2
Analysis of protein levels of Pol3 and Rfa2 in the wild-type and tetracycline downregulatable strains.
(A) Effect of downregulation of Pol3. (B) Effect of downregulation of Rfa2. Pol3 and Rfa2 were c-Myc tagged at the C-terminus in the wild-type, TET-POL3 and TET-RFA2 strains respectively. Proteins were extracted with (+) or without (−) treatment with doxycycline. Pol3 and Rfa2 were detected by Western blot with anti-c-MYC antibody. The protein levels were compared against Pgk1 levels (detected with anti-Pgk1 specific antibody) which acted as the loading control. Upon treatment with doxycycline, Pol3 expression was lowered 10 fold (average of 9, 10 and 12) and Rfa2 expression was lowered 4-fold (average of 4.1, 4.4 and 3.7).