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Nucleic Acids Res. 2017 Sep 6;45(15):8806-8821. doi: 10.1093/nar/gkx491.

Mediator binds to boundaries of chromosomal interaction domains and to proteins involved in DNA looping, RNA metabolism, chromatin remodeling, and actin assembly.

Author information

1
Division of Developmental Biology, Eunice Kennedy Shriver National Institute for Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
2
Department of Biochemistry and Molecular Biology, Penn State College of Medicine, Hershey, PA 17033, USA.
3
Department of Medical Biochemistry and Biophysics Umeå University, 901 87 Umeå, Sweden.
4
Department of Physics and Astronomy, Rutgers University, Piscataway, NJ 08854, USA.
5
Center for Quantitative Biology, Rutgers University, Piscataway, NJ 08854, USA.

Abstract

Mediator is a multi-unit molecular complex that plays a key role in transferring signals from transcriptional regulators to RNA polymerase II in eukaryotes. We have combined biochemical purification of the Saccharomyces cerevisiae Mediator from chromatin with chromatin immunoprecipitation in order to reveal Mediator occupancy on DNA genome-wide, and to identify proteins interacting specifically with Mediator on the chromatin template. Tandem mass spectrometry of proteins in immunoprecipitates of mediator complexes revealed specific interactions between Mediator and the RSC, Arp2/Arp3, CPF, CF 1A and Lsm complexes in chromatin. These factors are primarily involved in chromatin remodeling, actin assembly, mRNA 3'-end processing, gene looping and mRNA decay, but they have also been shown to enter the nucleus and participate in Pol II transcription. Moreover, we have found that Mediator, in addition to binding Pol II promoters, occupies chromosomal interacting domain (CID) boundaries and that Mediator in chromatin associates with proteins that have been shown to interact with CID boundaries, such as Sth1, Ssu72 and histone H4. This suggests that Mediator plays a significant role in higher-order genome organization.

PMID:
28575439
PMCID:
PMC5587782
DOI:
10.1093/nar/gkx491
[Indexed for MEDLINE]
Free PMC Article

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