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Altered Mediator dynamics during heat shock in budding yeast

Debasish Sarkar, Z. Iris Zhu, Emily Paul, David Landsman, View ORCID ProfileRandall H. Morse
doi: https://doi.org/10.1101/2020.08.25.267088
Debasish Sarkar
1Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509
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Z. Iris Zhu
3Computational Biology Branch, National Center for Biotechnology Information, National Library of Medicine, NIH, Bethesda, MD 20814
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Emily Paul
1Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509
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David Landsman
3Computational Biology Branch, National Center for Biotechnology Information, National Library of Medicine, NIH, Bethesda, MD 20814
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Randall H. Morse
1Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509
2Department of Biomedical Sciences, University at Albany School of Public Health, Albany, New York, 12201-0509
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  • ORCID record for Randall H. Morse
  • For correspondence: randall.morse@health.ny.gov
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Abstract

The Mediator complex is central to transcription by RNA polymerase II (Pol II) in eukaryotes. In yeast, Mediator is recruited by activators via its tail module and then facilitates assembly of the pre-initiation complex (PIC), including Pol II, setting the stage for productive transcription. Mediator occupies proximal promoter regions only transiently prior to Pol II escape; interruption of the transcription cycle by inactivation or depletion of Kin28 inhibits Pol II escape and stabilizes Mediator occupancy at promoters. However, whether Mediator occupancy and dynamics differ for gene cohorts induced by stress or alternative growth conditions has not been examined on a genome-wide scale. Here we investigate Mediator occupancy following heat shock or CdCl2 induction, with or without depletion of Kin28. We find that Pol II occupancy exhibits similar dependence on Mediator under normal and heat shock conditions; however, Mediator occupancy does not increase upon Kin28 depletion at most genes active during heat shock, indicating altered dynamics. Furthermore, Mediator occupancy persists at genes repressed by heat shock or CdCl2 induction and exhibits peaks upstream of the proximal promoter whether or not Kin28 is depleted, suggesting that Mediator is recruited by activators but is unable to engage PIC components at these repressed targets. Finally, we show a reduced dependence on PIC components for Mediator occupancy at promoters after heat shock, further supporting an altered dynamics or stronger engagement with activators under these conditions.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. This article is a US Government work. It is not subject to copyright under 17 USC 105 and is also made available for use under a CC0 license.
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Posted August 26, 2020.
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Altered Mediator dynamics during heat shock in budding yeast
Debasish Sarkar, Z. Iris Zhu, Emily Paul, David Landsman, Randall H. Morse
bioRxiv 2020.08.25.267088; doi: https://doi.org/10.1101/2020.08.25.267088
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Altered Mediator dynamics during heat shock in budding yeast
Debasish Sarkar, Z. Iris Zhu, Emily Paul, David Landsman, Randall H. Morse
bioRxiv 2020.08.25.267088; doi: https://doi.org/10.1101/2020.08.25.267088

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