Article Figures & Data
Figures
- Fig. 1.
Gcn5 is required for HO expression. (A) Effects of gcn5 disruption on the mRNA levels of theHO, PHO2, and SWI5 genes. Total RNA was extracted from FY120 (GCN5) and JJY54 (gcn5::hisG) grown in YEPD medium to mid-log phase. ACT1 mRNA was used as a control. (B) Genetic relationships between SWI5 and GCN5. β-Galactosidase activity was measured in strains carrying anHO-lacZ reporter gene integrated in the chromosome at theHO locus. The strains used were JJY12 (wild type [wt]), JJY28 (gcn5::hisG), JJY13 (swi5::hisG), and JJY60 (gcn5::hisG swi5::hisG). Values are averages of three independent measurements with less than 10% deviation.
- Fig. 2.
A deletion of the RPD3 gene partially suppresses the defects caused by a disruption of the GCN5gene. Cultures of JJY1 (wild type [wt]), JJY64 (rpd3Δ::LEU2), JJY28 (gcn5::hisG), and JJY65 (rpd3Δ::LEU2 gcn5::hisG) cells (approximately 5 × 106/ml) were spotted in 10-fold serial dilutions on medium lacking histidine (SD-HIS) and on medium lacking histidine and containing 10 mM 3-AT. Plates were incubated at 30°C for 3 days. The same cultures were used to measure β-galactosidase activity (in Miller units). Values are averages of three independent measurements with less than 10% deviation.
- Fig. 3.
Deletion of the SIN1 gene alleviates the defects associated with disruption of the GCN5 gene. Cultures of JJY12 (wild type [wt]), JJY36 (sin1Δ::TRP1), JJY28 (gcn5::hisG), and JJY45 (sin1Δ::TRP1 gcn5::hisG) cells (approximately 5 × 106/ml) were spotted in 10-fold serial dilutions on medium lacking histidine (SD-HIS) and on medium lacking histidine and containing 10 mM 3-AT. Plates were incubated at 30°C for 3 days. The same cultures were used to measure β-galactosidase activity (in Miller units). Values are averages of three independent measurements with less than 10% deviation.
- Fig. 4.
Histone sin mutations suppressgcn5 defects. (A) Cultures of JJY12 (wild type [wt]), JJY28 (gcn5::hisG), JJY41 (hhf2-7 gcn5::hisG), JJY42 (hhf2-8 gcn5::hisG), JJY43 (hhf2-13 gcn5::hisG), and JJY44 (sin2-1 gcn5::hisG) cells (approximately 5 × 106/ml) were spotted in 10-fold serial dilutions on medium lacking histidine (SD-HIS) and on medium lacking histidine and containing 10 mM 3-AT. Plates were incubated at 30°C for 3 days. The same cultures were used to measure β-galactosidase activity (in Miller units). Values are averages of three independent measurements with less than 10% deviation. (B) Effects of rpd3 deletion on the suppression of gcn5 defects by histone sinmutations and sin1 mutations. The strains used were JJY12, JJY28, and JJY41 through JJY44 (all as described for panel A), as well as JJY65 (rpd3Δ::LEU2 gcn5::hisG), JJY72 (hhf2-7 rpd3Δ::LEU2 gcn5::hisG), JJY73 (hhf2-8 rpd3Δ::LEU2 gcn5::hisG), JJY74 (hhf2-13 rpd3Δ::LEU2 gcn5::hisG), and JJY75 (sin2-1 rpd3Δ::LEU2 gcn5::hisG). Values are averages of three independent measurements with less than 10% deviation.
Tables
- Table 1.
Yeast strains used in this study
Strain Genotype FY120 MAT a ura3-52 leu2Δ1 his4-912δ Lys2-128δ RT238 MATα ura3-52 leu2Δ1 his3 trp1 HO-lacZ JJY12 MATα ura3-52 leu2 Δ1 trp1 lys2-128δ HO-lacZ JJY13 Same as JJY12, plusswi5::hisG JJY28 Same as JJY12, plusgcn5::hisG JJY36 Same as JJY12, plussin1Δ::TRP1 JJY42 Same as JJY28, plushhf2-8 JJY43 Same as JJY28, plus hhf2-13 JJY44 Same as JJY28, plus sin2-1 JJY45 Same as JJY28, plus sin1Δ::TRP1 JJY54 Same as FY120, plus gcn5::hisG JJY60 Same as JJY28, plusswi5::hisG JJY64 Same as JJY12, plusrpd3Δ::LEU2 JJY65 Same as JJY28, plusrpd3Δ::LEU2 JJY72 Same as JJY41, plusrpd3Δ::LEU2 JJY73 Same as JJY42, plusrpd3Δ::LEU2 JJY74 Same as JJY43, plusrpd3Δ::LEU2 JJY75 Same as JJY44, plusrpd3Δ::LEU2
