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A DNA-tethered cleavage probe reveals the path for promoter DNA in the yeast preinitiation complex.

Miller, Gail and Hahn, Steven (2006) A DNA-tethered cleavage probe reveals the path for promoter DNA in the yeast preinitiation complex. Nature structural & molecular biology, 13 (7). pp. 603-610. ISSN 1545-9993

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Article URL: http://www.nature.com/nsmb/journal/v13/n7/abs/nsmb...

Abstract

To directly map the position of promoter DNA within the RNA polymerase II (Pol II) transcription preinitiation complex (PIC), FeBABE was tethered to specific sites within the HIS4 promoter and used to map exposed surfaces of Pol II and the general transcription factors in proximity to DNA. Our results distinguish between previously proposed models for PIC structure and demonstrate that downstream promoter DNA is positioned over the central cleft of Pol II, with DNA upstream of TATA extending toward the Pol II subunit Rpb3. Also mapped were segments of TFIIB, TFIIE, TFIIF and TFIIH in proximity to promoter DNA. DNA downstream of the transcription bubble maps to a path between the two helicase subdomains of the TFIIH subunit Rad25 (also called XPB). Together, our results show how the general factors and Pol II converge on promoter DNA within the PIC.

Item Type: Article
Additional Information: This article is available to subscribers only via the URL above.
DOI: 10.1038/nsmb1117
PubMed ID: 16819517
NIHMSID: NIHMS57561
PMCID: PMC2483788
Grant Numbers: R01 GM053451-12
Keywords or MeSH Headings: Base Sequence; DNA Helicases/chemistry/genetics/metabolism; DNA Polymerase II/chemistry/genetics; DNA, Fungal/chemistry/genetics; Hydroxyl Radical; Models, Molecular; Nucleic Acid Conformation; Promoter Regions (Genetics); Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae Proteins/chemistry/genetics/metabolism; Transcription Factor TFIIH/chemistry/genetics; Transcription, Genetic;
Subjects: Molecules > Proteins > Transcription factors
Cellular and Organismal Processes > Genetic processes > Transcription
Depositing User: Library Staff
Date Deposited: 14 Aug 2008 22:04
Last Modified: 14 Feb 2012 14:42
URI: http://authors.fhcrc.org/id/eprint/8

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