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TitleMolecular basis of transcription initiation in Archaea.
Journal, issue, pagesTranscription, Vol. 1, Issue 2, Page 103-111, Year 2010
Publish dateJun 13, 2012
AuthorsSacha De Carlo / Shih-Chieh Lin / Dylan J Taatjes / Andreas Hoenger /
PubMed AbstractCompared with eukaryotes, the archaeal transcription initiation machinery-commonly known as the Pre-Initiation Complex-is relatively simple. The archaeal PIC consists of the TFIIB ortholog TFB, TBP, ...Compared with eukaryotes, the archaeal transcription initiation machinery-commonly known as the Pre-Initiation Complex-is relatively simple. The archaeal PIC consists of the TFIIB ortholog TFB, TBP, and an 11-subunit RNA polymerase (RNAP). The relatively small size of the entire archaeal PIC makes it amenable to structural analysis. Using purified RNAP, TFB, and TBP from the thermophile Pyrococcus furiosus, we assembled the biochemically active PIC at 65ºC. The intact archaeal PIC was isolated by implementing a cross-linking technique followed by size-exclusion chromatography, and the structure of this 440 kDa assembly was determined using electron microscopy and single-particle reconstruction techniques. Combining difference maps with crystal structure docking of various sub-domains, TBP and TFB were localized within the macromolecular PIC. TBP/TFB assemble near the large RpoB subunit and the RpoD/L "foot" domain behind the RNAP central cleft. This location mimics that of yeast TBP and TFIIB in complex with yeast RNAP II. Collectively, these results define the structural organization of the archaeal transcription machinery and suggest a conserved core PIC architecture.
External linksTranscription / PubMed:21326901 / PubMed Central
MethodsEM (single particle)
Resolution18.0 - 25.0 Å
Structure data

EMDB-5024:
Cryo-EM structure of Pyrococcus furiosus in apo-state
Method: EM (single particle) / Resolution: 18.0 Å

EMDB-5025:
Cryo-EM structure of Pyrococcus furiosus pre-initiation complex
Method: EM (single particle) / Resolution: 25.0 Å

Source
  • unidentified (others)

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