Fig. 5
From: Identification and characterization of sugar-regulated promoters in Chaetomium thermophilum
The xylose-inducible PXDH promoter controls a dominant-negative mutant eliciting a 60S export defect. a Cartoon showing the reporter assay for monitoring an inducible localization defect of the large 60S subunit using L25-YFP. We constitutively expressed L25-YFP using the hph1 selection marker and introduced additional plasmids that allow overexpressing of the ribosome biogenesis factor pA-Rsa4 (PXDH-RSA4) or the mutated protein pA-rsa4 E117D (PXDH-rsa4) under the control of PXDH, using the erg1 selection marker. Based on the rsa4 E117D mutant from yeast, a dominant-negative growth phenotype and the accumulation of L25-YFP in the nucleoplasm is expected upon induction of the mutant protein. b Immunoblotting shows the controlled expression of pA-rsa4 E117D on xylose-containing medium at the indicated time points. The uncropped blot is shown in Related file 3. c L25-YFP localization in RSA4- and rsa4 E117D-expressing strains under the control of PXDH. L25-YFP strains expressing pA-RSA4 (RSA4) or pA-RSA4 E117D (rsa4) under the control of PXDH were grown for 16 h in glucose-containing medium and then shifted to fresh glucose- or xylose-containing medium for another 4 h. Images were acquired in the DIC and YFP channels. Unprocessed blots/gels are presented in Supplementary Fig. 8