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. 2022 Jul 1;9:896215. doi: 10.3389/fmolb.2022.896215

FIGURE 4.

FIGURE 4

Identification of host rhf and rtt mutants that are dependent on the low activity or architecture of the U3 promoter. (A) A semi-quantitative assay comparing retromobility of Ty1his3AI on the identical CEN-plasmid and driven from the P TEF1 , U3 or P PSP2 promoter. His+ Leu+ papillation (indicative of Ty1his3AI retromobility) of three independent transformants of the wild-type strain carrying the LEU2-marked CEN-plasmid with P PSP2 -Ty1his3AI, P TEF1 -Ty1his3AI or U3-Ty1his3AI, as indicated, following growth on YPD agar at 20°C to induce retrotransposition. (B) Graph showing the mean retromobility frequency (MRF) in six independent transformants of the CEN-P PSP2 -Ty1his3AI plasmid in the wild-type (light blue bars) or congenic med15∆::URA3 (dark blue bars) strain bearing a chromosomal Ty1kanMXAI element. Solid bars- frequency of G418R reversion (indicative of Ty1kanMXAI retromobility); checked bars-frequency of His+ reversion (indicative of Ty1his3AI retromobility). Error bars; standard error. The absence of error bars on dark blue or dark-blue-checked bars indicates that the value is an estimated maximum retromobility frequency because zero G418R or His+ colonies were observed in cultures of six independent transformants of the med15∆ strain. (C) A SC-Leu-His plate from the P PSP2 -Ty1his3AI screen showing relative levels of His+ Leu+ papillation (indicative of Ty1his3AI retromobility) in transformants of the wild-type strain, yhr130C∆ (an rhf mutant with no change in His+ Leu+ papillation relative to the wild-type strain), ygl214w∆ (an rhf mutant with decreased His+ Leu+ papillation) and yel008w∆ (an rtt mutant with increased His+ Leu+ papillation) mutants. (D) Lists of U3-low activity dependent regulators (U3laDR) in red and U3-architecture dependent regulators (U3arDR) in blue, as identified by the P PSP2 -Ty1his3AI screen. Bold type indicates RTT genes; regular type indicates RHF genes. MED15, indicated in black, is a control.

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