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Mediator12 and 13 act as conditional anti-silencing factors in Arabidopsis [ChIP-seq]

ABSTRACT: Mediator is a multi-protein complex which facilitates the initial steps of gene transcription. Mediator 12 (MED12) and MED13 are components of a module that reversibly associates with the core Mediator complex, and both positively and negatively regulates gene expression. Here, in an Arabidopsis mutant screen for factors that can bypass repressive epigenetic marks, we identified MED12 and MED13 as anti-silencing factors. Both are preferentially required for the expression of genes in the absence of H3K4me3, an activating chromatin mark. Thus, MED12 and MED13 respond to specific epigenetic states and can act as conditional positive gene regulators in Arabidopsis.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE123562 | GEO | 2020/12/31

REPOSITORIES: GEO

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Mediator12 and 13 act as conditional anti-silencing factors in Arabidopsis [RNA-seq]

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The characterization of Mediator12 and 13 as conditional positive gene regulators in Arabidopsis [RNA-seq]

Project description:Mediator is a multi-protein complex which facilitates the initial steps of gene transcription. Mediator 12 (MED12) and MED13 are components of a module that reversibly associates with the core Mediator complex, and both positively and negatively regulates gene expression. Here, in an Arabidopsis mutant screen for factors required for morc1 de-repressed gene expression, we identified MED12 and MED13 as positive gene regulators, both of which contribute broadly to morc1 de-repressed gene expression. Both MED12 and MED13 are preferentially required for the expression of genes lacking active chromatin marks, a chromatin signature shared with morc1 re-activated loci. We further discovered that MED12 tends to interact with genes that are responsive to environmental stimuli, including light and radiation. We demonstrate that the light-induced transient gene expression depends on MED12, and is accompanied by a concomitant increase in MED12 enrichment during induction. In contrast, the steady-state expression level of these genes showed little dependence on MED12, suggesting that MED12 is primarily required to aid the expression of genes in transition from non-active to active states.

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Project description:The Mediator complex plays a pivotal role in facilitating RNA polymerase II-dependent transcription in eukaryotes. Within this complex, the CDK8 kinase module (CKM), comprising CDK8, Cyclin C (CycC), Med12, and Med13, serves as a dissociable subcomplex that modulates the activity of the small Mediator complex. Genetic studies in Drosophila have revealed distinct phenotypes associated with mutations in CKM subunits, yet the underlying mechanism has remained unknown. Using Drosophila as a model, we generated transgenic strains to individually or simultaneously deplete the four CKM subunits in all possible combinations, uncovering unique phenotypes in the eyes and wings. Depletion of CDK8-CycC enhanced E2F1 target gene expression and promoted cell-cycle progression, whereas Med12-Med13 depletion had no significant impact on these processes. Conversely, depleting Med12-Med13 altered the expression of ribosomal protein genes and fibrillarin, reduced nascent protein synthesis, indicating a severe reduction in ribosome biogenesis and cellular growth compared to the loss of CDK8-CycC. These findings reveal distinct in vivo roles for CKM subunits, with Med12-Med13 disruption having a more pronounced effect on ribosome biogenesis and protein synthesis .

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