Limits...
Expression profiling of S. pombe acetyltransferase mutants identifies redundant pathways of gene regulation.

Nugent RL, Johnsson A, Fleharty B, Gogol M, Xue-Franzén Y, Seidel C, Wright AP, Forsburg SL - BMC Genomics (2010)

Bottom Line: Comparison of phenotypes and expression profiles in single, double and triple mutants indicate that these HAT enzymes have overlapping functions.Consistent with this, overlapping specificity in histone H3 acetylation is observed.However, there is no evidence for overlap with another HAT enzyme, encoded by the essential mst1+ gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular and Computational Biology Section, University of Southern California, Los Angeles, California 90089-2910, USA.

ABSTRACT

Background: Histone acetyltransferase enzymes (HATs) are implicated in regulation of transcription. HATs from different families may overlap in target and substrate specificity.

Results: We isolated the elp3+ gene encoding the histone acetyltransferase subunit of the Elongator complex in fission yeast and characterized the phenotype of an Deltaelp3 mutant. We examined genetic interactions between Deltaelp3 and two other HAT mutants, Deltamst2 and Deltagcn5 and used whole genome microarray analysis to analyze their effects on gene expression.

Conclusions: Comparison of phenotypes and expression profiles in single, double and triple mutants indicate that these HAT enzymes have overlapping functions. Consistent with this, overlapping specificity in histone H3 acetylation is observed. However, there is no evidence for overlap with another HAT enzyme, encoded by the essential mst1+ gene.

Show MeSH

Related in: MedlinePlus

Characterization of mutant phenotypes . A: Δelp3 morphology. Wild-type (FY368) and Δelp3 (FY3851)cells were asynchronously grown at 32°C in YES. Cells were fixed and stained with DAPI. The scale bar represents 10 microns. B: Δelp3 mutants have a growth lag. Representative growth curves for wild type (triangle) (FY368) and Δelp3 (circle) (FY3851) cells in rich media (YES) at 30°C. The x-axis time scale indicates hours after inoculation that stared at a cell concentration of 4 × 105 cells/ml. The y-axis indicates number of cells/ml. C: Comparison of growth rates on YES at different temperatures and with different levels of salt. Cells were grown to exponential phase, serially diluted 5× and compared for growth on rich media after 2 or 3 days at 30°C. and 36°C, or onYES supplemented with indicated salt and grown for 2 to 4 days at 30°C. Strain list: wild-type (FY368), Δelp3(FY3851), Δmst2 (FY1890), Δgcn5 Δmst2(Hu990), Δgcn5 Δelp3 (FY3847), Δmst2 Δelp3 (FY3850), Δgcn5 Δelp3 Δmst2 (3854) D: Comparison of drug sensitivity. Exponentially growing cells were diluted 5-fold on YES plates with the indicated drugs and grown 2-3 days at 32°C. Strain list: wild-type (FY261), Δswi6 (FY1584), Δrad3 (FY1104), Δelp3 (FY 3851), Δgcn5 (FY2943), Δmst2 (FY1890), Δgcn5 Δelp3 (FY3847), Δgcn5 Δmst2(FY3361), Δmst2 Δelp3 (FY3850), Δgcn5 Δelp3 Δmst2 (3854).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2823694&req=5

Figure 1: Characterization of mutant phenotypes . A: Δelp3 morphology. Wild-type (FY368) and Δelp3 (FY3851)cells were asynchronously grown at 32°C in YES. Cells were fixed and stained with DAPI. The scale bar represents 10 microns. B: Δelp3 mutants have a growth lag. Representative growth curves for wild type (triangle) (FY368) and Δelp3 (circle) (FY3851) cells in rich media (YES) at 30°C. The x-axis time scale indicates hours after inoculation that stared at a cell concentration of 4 × 105 cells/ml. The y-axis indicates number of cells/ml. C: Comparison of growth rates on YES at different temperatures and with different levels of salt. Cells were grown to exponential phase, serially diluted 5× and compared for growth on rich media after 2 or 3 days at 30°C. and 36°C, or onYES supplemented with indicated salt and grown for 2 to 4 days at 30°C. Strain list: wild-type (FY368), Δelp3(FY3851), Δmst2 (FY1890), Δgcn5 Δmst2(Hu990), Δgcn5 Δelp3 (FY3847), Δmst2 Δelp3 (FY3850), Δgcn5 Δelp3 Δmst2 (3854) D: Comparison of drug sensitivity. Exponentially growing cells were diluted 5-fold on YES plates with the indicated drugs and grown 2-3 days at 32°C. Strain list: wild-type (FY261), Δswi6 (FY1584), Δrad3 (FY1104), Δelp3 (FY 3851), Δgcn5 (FY2943), Δmst2 (FY1890), Δgcn5 Δelp3 (FY3847), Δgcn5 Δmst2(FY3361), Δmst2 Δelp3 (FY3850), Δgcn5 Δelp3 Δmst2 (3854).

Mentions: In order to compare the roles of different histone acetyltransferases on transcription in fission yeast, we first characterized the disruption of the histone acetyltransferase gene elp3+, the likely orthologue of the ScELP3 subunit of Elongator complex [21]. We found that S. pombe elp3+ is a non-essential gene, as is its budding yeast orthologue [30]. However, Δelp3 mutant cells have elongated morphology compared to wild type, indicative of cell cycle delay (Fig. 1A). The mean length of mononucleate cells is 6.7 μm in wild type, and 10.7 μm in Δelp3. For binucleate cells, wild type averages 9.8 μm compared to 15.9 μm in Δelp3. Thus Δelp3 is approximately 60% larger than wild type. Also as in budding yeast [30], fission yeast Δelp3 grows slowly due to a prolonged lag phase (Fig. 1B). Although the growth rate of Δelp3 cells once they have reached exponential phase is similar to wild type, they appear to enter stationary phase at a lower cell density. Given the presumed role of Elp3 as part of the Elongator complex, we presume that this reflects defects in one or more growth-associated transcription program.


Expression profiling of S. pombe acetyltransferase mutants identifies redundant pathways of gene regulation.

Nugent RL, Johnsson A, Fleharty B, Gogol M, Xue-Franzén Y, Seidel C, Wright AP, Forsburg SL - BMC Genomics (2010)

Characterization of mutant phenotypes . A: Δelp3 morphology. Wild-type (FY368) and Δelp3 (FY3851)cells were asynchronously grown at 32°C in YES. Cells were fixed and stained with DAPI. The scale bar represents 10 microns. B: Δelp3 mutants have a growth lag. Representative growth curves for wild type (triangle) (FY368) and Δelp3 (circle) (FY3851) cells in rich media (YES) at 30°C. The x-axis time scale indicates hours after inoculation that stared at a cell concentration of 4 × 105 cells/ml. The y-axis indicates number of cells/ml. C: Comparison of growth rates on YES at different temperatures and with different levels of salt. Cells were grown to exponential phase, serially diluted 5× and compared for growth on rich media after 2 or 3 days at 30°C. and 36°C, or onYES supplemented with indicated salt and grown for 2 to 4 days at 30°C. Strain list: wild-type (FY368), Δelp3(FY3851), Δmst2 (FY1890), Δgcn5 Δmst2(Hu990), Δgcn5 Δelp3 (FY3847), Δmst2 Δelp3 (FY3850), Δgcn5 Δelp3 Δmst2 (3854) D: Comparison of drug sensitivity. Exponentially growing cells were diluted 5-fold on YES plates with the indicated drugs and grown 2-3 days at 32°C. Strain list: wild-type (FY261), Δswi6 (FY1584), Δrad3 (FY1104), Δelp3 (FY 3851), Δgcn5 (FY2943), Δmst2 (FY1890), Δgcn5 Δelp3 (FY3847), Δgcn5 Δmst2(FY3361), Δmst2 Δelp3 (FY3850), Δgcn5 Δelp3 Δmst2 (3854).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2823694&req=5

Figure 1: Characterization of mutant phenotypes . A: Δelp3 morphology. Wild-type (FY368) and Δelp3 (FY3851)cells were asynchronously grown at 32°C in YES. Cells were fixed and stained with DAPI. The scale bar represents 10 microns. B: Δelp3 mutants have a growth lag. Representative growth curves for wild type (triangle) (FY368) and Δelp3 (circle) (FY3851) cells in rich media (YES) at 30°C. The x-axis time scale indicates hours after inoculation that stared at a cell concentration of 4 × 105 cells/ml. The y-axis indicates number of cells/ml. C: Comparison of growth rates on YES at different temperatures and with different levels of salt. Cells were grown to exponential phase, serially diluted 5× and compared for growth on rich media after 2 or 3 days at 30°C. and 36°C, or onYES supplemented with indicated salt and grown for 2 to 4 days at 30°C. Strain list: wild-type (FY368), Δelp3(FY3851), Δmst2 (FY1890), Δgcn5 Δmst2(Hu990), Δgcn5 Δelp3 (FY3847), Δmst2 Δelp3 (FY3850), Δgcn5 Δelp3 Δmst2 (3854) D: Comparison of drug sensitivity. Exponentially growing cells were diluted 5-fold on YES plates with the indicated drugs and grown 2-3 days at 32°C. Strain list: wild-type (FY261), Δswi6 (FY1584), Δrad3 (FY1104), Δelp3 (FY 3851), Δgcn5 (FY2943), Δmst2 (FY1890), Δgcn5 Δelp3 (FY3847), Δgcn5 Δmst2(FY3361), Δmst2 Δelp3 (FY3850), Δgcn5 Δelp3 Δmst2 (3854).
Mentions: In order to compare the roles of different histone acetyltransferases on transcription in fission yeast, we first characterized the disruption of the histone acetyltransferase gene elp3+, the likely orthologue of the ScELP3 subunit of Elongator complex [21]. We found that S. pombe elp3+ is a non-essential gene, as is its budding yeast orthologue [30]. However, Δelp3 mutant cells have elongated morphology compared to wild type, indicative of cell cycle delay (Fig. 1A). The mean length of mononucleate cells is 6.7 μm in wild type, and 10.7 μm in Δelp3. For binucleate cells, wild type averages 9.8 μm compared to 15.9 μm in Δelp3. Thus Δelp3 is approximately 60% larger than wild type. Also as in budding yeast [30], fission yeast Δelp3 grows slowly due to a prolonged lag phase (Fig. 1B). Although the growth rate of Δelp3 cells once they have reached exponential phase is similar to wild type, they appear to enter stationary phase at a lower cell density. Given the presumed role of Elp3 as part of the Elongator complex, we presume that this reflects defects in one or more growth-associated transcription program.

Bottom Line: Comparison of phenotypes and expression profiles in single, double and triple mutants indicate that these HAT enzymes have overlapping functions.Consistent with this, overlapping specificity in histone H3 acetylation is observed.However, there is no evidence for overlap with another HAT enzyme, encoded by the essential mst1+ gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular and Computational Biology Section, University of Southern California, Los Angeles, California 90089-2910, USA.

ABSTRACT

Background: Histone acetyltransferase enzymes (HATs) are implicated in regulation of transcription. HATs from different families may overlap in target and substrate specificity.

Results: We isolated the elp3+ gene encoding the histone acetyltransferase subunit of the Elongator complex in fission yeast and characterized the phenotype of an Deltaelp3 mutant. We examined genetic interactions between Deltaelp3 and two other HAT mutants, Deltamst2 and Deltagcn5 and used whole genome microarray analysis to analyze their effects on gene expression.

Conclusions: Comparison of phenotypes and expression profiles in single, double and triple mutants indicate that these HAT enzymes have overlapping functions. Consistent with this, overlapping specificity in histone H3 acetylation is observed. However, there is no evidence for overlap with another HAT enzyme, encoded by the essential mst1+ gene.

Show MeSH
Related in: MedlinePlus