The Initiation Factor TFE and the Elongation Factor Spt4/5 Compete for the RNAP Clamp during Transcription Initiation and Elongation

• Published in: Molecular cell Vol. 43; no. 2; pp. 263 - 274
• Main Authors: Grohmann, Dina, Nagy, Julia, Chakraborty, Anirban, Klose, Daniel, Fielden, Daniel, Ebright, Richard H., Michaelis, Jens, Werner, Finn
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 22-07-2011; Cell Press
• Subjects: Amino Acid Sequence; Archaea - genetics; Archaea - metabolism; Archaeal Proteins - chemistry; Archaeal Proteins - genetics; Archaeal Proteins - metabolism; Base Sequence; Binding Sites; Chromosomal Proteins, Non-Histone - chemistry; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; DNA; DNA-Directed RNA Polymerases - metabolism; Fluorescence Resonance Energy Transfer; Methanocaldococcus jannaschii; Models, Molecular; Molecular Sequence Data; promoter regions; Protein Conformation; Sulfolobus solfataricus - metabolism; transcription (genetics); transcription factors; Transcription, Genetic; Transcriptional Elongation Factors - chemistry; Transcriptional Elongation Factors - genetics; Transcriptional Elongation Factors - metabolism
• ISSN: 1097-2765; 1097-4164
• DOI: 10.1016/j.molcel.2011.05.030

TFIIE and the archaeal homolog TFE enhance DNA strand separation of eukaryotic RNAPII and the archaeal RNAP during transcription initiation by an unknown mechanism. We have developed a fluorescently labeled recombinant M. jannaschii RNAP system to probe the archaeal transcription initiation complex, consisting of promoter DNA, TBP, TFB, TFE, and RNAP. We have localized the position of the TFE winged helix (WH) and Zinc ribbon (ZR) domains on the RNAP using single-molecule FRET. The interaction sites of the TFE WH domain and the transcription elongation factor Spt4/5 overlap, and both factors compete for RNAP binding. Binding of Spt4/5 to RNAP represses promoter-directed transcription in the absence of TFE, which alleviates this effect by displacing Spt4/5 from RNAP. During elongation, Spt4/5 can displace TFE from the RNAP elongation complex and stimulate processivity. Our results identify the RNAP “clamp” region as a regulatory hot spot for both transcription initiation and transcription elongation.
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► The RNAP clamp coiled coil and RNAP stalk are required for TFE binding and activity ► The elongation factor Spt4/5 can inhibit PIC formation and transcription initiation ► TFE efficiently prevents inhibition of transcription initiation by Spt4/5 ► Spt4/5 displaces TFE from the TEC