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The Transcriptional Regulator Ume6 is a Major Driver of Gene Expression during Gametogenesis in Budding Yeast

ABSTRACT: The Transcriptional Regulator Ume6 is a Major Driver of Gene Expression during Gametogenesis in Budding Yeast

PROVIDER: PRJNA941371 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR23717400.fastq.gz	Fastqsanger.gz
	SRR23717500.fastq.gz	Fastqsanger.gz
	SRR23717401.fastq.gz	Fastqsanger.gz
	SRR23717501.fastq.gz	Fastqsanger.gz
	SRR23717402.fastq.gz	Fastqsanger.gz

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The Transcriptional Regulator Ume6 is a Major Driver of Gene Expression during Gametogenesis in Budding Yeast

Project description:Early meiotic genes (EMGs) encompass an important subset within the developmental program of gametogenesis. Regulation of EMGs is achieved through the recognition of a common regulatory element in their promoters by the transcription factor Ume6. Proper meiotic execution requires the timely expression of EMGs, which is achieved through the adoption of the meiotic regulator Ime1 by Ume6. While it is known that Ime1’s binding to Ume6 promotes EMG expression, the mechanism of EMG activation remains unclear. Two competing models have been proposed: (1) Ime1 converts Ume6 to a coactivator complex, and (2) Ime1 binding targets Ume6 for degradation. Using a combination of tools including meiotic synchronization, temporally controlled protein depletion, protein-protein interaction engineering, and transcriptomics, here we address this conundrum. We identify 160 genes that are directly regulated by Ime1-Ume6, including UME6 itself. Removal of Ume6 shortly before meiotic entry is detrimental to EMG activation and gamete formation, whereas tethering of Ume6 to a heterologous activation domain is sufficient to trigger EMG expression and to generate viable gametes in the absence of Ime1. We conclude that Ime1-Ume6 functions as a coactivator complex. While Ume6 is indispensable for EMGs expression, Ime1 primarily serves as an evolutionarily tuned activation domain for Ume6.

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Transcription profiling of S. cerevisiae (budding yeast) ume6 deletion strains vs wild-type in different media to determine the complement of Ume6 dependent genes during mitosis and meiosis

Project description:To determine the complement of Ume6-dependent genes expressed during mitosis and/or meiosis in budding yeast we compared wild-type and<br>ume6 deletion strains using Yeast 2.0 high density oligonucleotide microarrays (GeneChips). Samples were analysed from cells growing in rich medium with fermentable (glucose) and non-fermentable (acetate) carbon sources and from cells undergoing meiosis and spore formation in sporulation medium. Expression data were combined with data from a genome-wide Ume6 DNA binding assay and Ume6-target site prediction to identify the most likely direct target genes of Ume6.

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Project description:Starvation in diploid budding yeast cells triggers a cell-fate program culminating in meiosis and spore formation. Transcription activation of early meiotic genes (EMGs) hinges on the transcription activator Ime1, its DNA-binding partner Ume6, and GSK-3 kinase Rim11. Phosphorylation of Ume6 by Rim11 is key for EMG activation. We report that Rim11 functions as the central signal integrator for controlling Ume6 phosphorylation and EMG transcription. In nutrient-rich conditions, PKA suppresses Rim11 levels, while TORC1 retains Rim11 in the cytoplasm. Inhibiting PKA and TORC1 induces Rim11 expression and nuclear localization. Remarkably, nuclear Rim11 is required, but not sufficient, for Rim11-dependent Ume6 phosphorylation. Additionally, Ime1 is an essential anchor protein for phosphorylating Ume6. Subsequently, Ume6-Ime1 coactivator complexes form, which drive EMG transcription. Our results demonstrate how varied signalling inputs (PKA/TORC1/Ime1) converge through Rim11 to regulate EMG expression and meiosis initiation. We posit that the signalling-regulatory network elucidated here generates robustness in cell-fate control.

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