Reconstitution of RNA Polymerase I Upstream Activating Factor and the Roles of Histones H3 and H4 in Complex Assembly
| Autor: | Weidong Cui, Nancy Walker-Kopp, Ashleigh J. Jackobel, Bruce A. Knutson, Marissa L Smith |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Transcriptional Activation
0301 basic medicine Saccharomyces cerevisiae Proteins Protein subunit Upstream activating factor Saccharomyces cerevisiae Biochemistry Histones Histone H4 03 medical and health sciences Histone H3 RNA Polymerase I Structural Biology Transcription (biology) Genetics Escherichia coli RNA polymerase I DNA Fungal Promoter Regions Genetic Molecular Biology biology Chemistry biology.organism_classification Recombinant Proteins Cell biology DNA-Binding Proteins 030104 developmental biology Histone biology.protein Biotechnology Transcription Factors |
| Zdroj: | Journal of Molecular Biology. 430:641-654 |
| ISSN: | 0022-2836 |
| DOI: | 10.1016/j.jmb.2018.01.003 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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