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1. Role of the pre-initiation complex in Mediator recruitment and dynamics

Author

Elisabeth R Knoll, Z Iris Zhu, Debasish Sarkar, David Landsman, and Randall H Morse

Subjects
mediator, transcription factors, TATA-binding protein, pre-initiation complex, yeast, ChIP-seq, Medicine, Science, Biology (General), QH301-705.5
Abstract

The Mediator complex stimulates the cooperative assembly of a pre-initiation complex (PIC) and recruitment of RNA Polymerase II (Pol II) for gene activation. The core Mediator complex is organized into head, middle, and tail modules, and in budding yeast (Saccharomyces cerevisiae), Mediator recruitment has generally been ascribed to sequence-specific activators engaging the tail module triad of Med2-Med3-Med15 at upstream activating sequences (UASs). We show that yeast lacking Med2-Med3-Med15 are viable and that Mediator and PolII are recruited to promoters genome-wide in these cells, albeit at reduced levels. To test whether Mediator might alternatively be recruited via interactions with the PIC, we examined Mediator association genome-wide after depleting PIC components. We found that depletion of Taf1, Rpb3, and TBP profoundly affected Mediator association at active gene promoters, with TBP being critical for transit of Mediator from UAS to promoter, while Pol II and Taf1 stabilize Mediator association at proximal promoters.

Published
2018
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2. Role of the pre-initiation complex in Mediator recruitment and dynamics

Author

Randall H. Morse, David Landsman, Debasish Sarkar, Elisabeth R. Knoll, and Z. Iris Zhu

Subjects
0301 basic medicine, Saccharomyces cerevisiae Proteins, Transcription, Genetic, QH301-705.5, Science, S. cerevisiae, RNA polymerase II, Saccharomyces cerevisiae, yeast, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Core mediator complex, Mediator, transcription factors, Gene Expression Regulation, Fungal, Biology (General), Promoter Regions, Genetic, Transcription factor, TATA-binding protein, Regulation of gene expression, TATA-Binding Protein Associated Factors, Mediator Complex, General Immunology and Microbiology, biology, Models, Genetic, Chemistry, General Neuroscience, Promoter, Genetics and Genomics, General Medicine, Chromosomes and Gene Expression, Cell biology, ChIP-seq, TAF1, 030104 developmental biology, pre-initiation complex, Mutation, biology.protein, Medicine, Transcription Factor TFIID, RNA Polymerase II, mediator, Research Article, Protein Binding
Abstract

The Mediator complex stimulates the cooperative assembly of a pre-initiation complex (PIC) and recruitment of RNA Polymerase II (Pol II) for gene activation. The core Mediator complex is organized into head, middle, and tail modules, and in budding yeast (Saccharomyces cerevisiae), Mediator recruitment has generally been ascribed to sequence-specific activators engaging the tail module triad of Med2-Med3-Med15 at upstream activating sequences (UASs). We show that yeast lacking Med2-Med3-Med15 are viable and that Mediator and PolII are recruited to promoters genome-wide in these cells, albeit at reduced levels. To test whether Mediator might alternatively be recruited via interactions with the PIC, we examined Mediator association genome-wide after depleting PIC components. We found that depletion of Taf1, Rpb3, and TBP profoundly affected Mediator association at active gene promoters, with TBP being critical for transit of Mediator from UAS to promoter, while Pol II and Taf1 stabilize Mediator association at proximal promoters.

Published
2018

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