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. 2001 Jul 10;98(15):8720–8725. doi: 10.1073/pnas.151240898

Figure 1.

Figure 1

pRB family members repress transcription in yeast. (A) Two reporters were used in the majority of repression assays. In the reporter used for growth assays, the UAS region from the HIS3 promoter is replaced with a region from the GAL1 promoter containing multiple Gal4p-binding sites (14). The reporter plasmid pRS314–17d80lacZdNeo (Lower) was used for all quantitation assays (19). (B) pRB family members repress transcription of the GAL1∷HIS3 reporter. Strain MaV103 was transformed with ARS-CEN plasmids expressing DB, DB-pRB, DB-p107, and DB-p130 and assayed for growth on either nonselective media (SD, −Leu) after 3 days or selective media (SD, −Leu, −His, 10 mM 3-AT) after 5 days. All growth assays were repeated several times with identical results. (C) Mutant alleles of pRB confer different levels of repression. Repression was assayed on SD, −Leu, −His, 10 mM 3-AT after 5 days. (D) pRB represses transcription in quantitative assays. All β-galactosidase assays were performed according to standard techniques. Activity units are presented as [A400/(A600 × time in minutes)] × 1,000. Data generated are the mean result and standard deviation of five independent cultures from one experiment including each DB construct. These results are representative of several independent experiments.