Reconstitution of RNA Polymerase I Upstream Activating Factor and the Roles of Histones H3 and H4 in Complex Assembly

Overview

Journal J Mol Biol

Publisher Elsevier

Specialties Microbiology
Molecular Biology

Date 2018 Jan 23

PMID 29357286

Citations 5

Authors

Marissa L Smith

Weidong Cui

Ashleigh J Jackobel

Nancy Walker-Kopp

Bruce A Knutson

Affiliations

Soon will be listed here.

Abstract

RNA polymerase I (Pol I) transcription in Saccharomyces cerevisiae requires four separate factors that recruit Pol I to the promoter to form a pre-initiation complex. Upstream Activating Factor (UAF) is one of two multi-subunit complexes that regulate pre-initiation complex formation by binding to the ribosomal DNA promoter and by stimulating recruitment of downstream Pol I factors. UAF is composed of Rrn9, Rrn5, Rrn10, Uaf30, and histones H3 and H4. We developed a recombinant Escherichia coli-based system to coexpress and purify transcriptionally active UAF complex and to investigate the importance of each subunit in complex formation. We found that no single subunit is required for UAF assembly, including histones H3 and H4. We also demonstrate that histone H3 is able to interact with each UAF-specific subunit, and show that there are at least two copies of histone H3 and one copy of H4 present in the complex. Together, our results provide a new model suggesting that UAF contains a hybrid H3-H4 tetramer-like subcomplex.

Citing Articles

Mechanism of RNA polymerase I selection by transcription factor UAF.

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Coupling Between Production of Ribosomal RNA and Maturation: Just at the Beginning.

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Molecular Topology of RNA Polymerase I Upstream Activation Factor.

Knutson B, Smith M, Belkevich A, Fakhouri A Mol Cell Biol. 2020; 40(13).

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