The RNA polymerase trigger loop functions in all three phases of the transcription cycle

The trigger loop (TL) forms a conserved element in the RNA polymerase active centre that functions in the elongation phase of transcription. Here, we show that the TL also functions in transcription initiation and termination. Using recombinant variants of RNA polymerase from Pyrococcus furiosus and...

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Bibliographic Details
Published in:Nucleic acids research 2013-08, Vol.41 (14), p.7048-7059
Main Authors: Fouqueau, Thomas, Zeller, Mirijam E, Cheung, Alan C, Cramer, Patrick, Thomm, Michael
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Archaeal Proteins - chemistry
Archaeal Proteins - genetics
Archaeal Proteins - metabolism
Conserved Sequence
DNA-Directed RNA Polymerases - chemistry
DNA-Directed RNA Polymerases - genetics
DNA-Directed RNA Polymerases - metabolism
Molecular Sequence Data
Mutation
Nucleic Acid Enzymes
Pyrococcus furiosus
Pyrococcus furiosus - enzymology
RNA - biosynthesis
RNA Cleavage
Sequence Homology, Amino Acid
Transcription Elongation, Genetic
Transcription Initiation, Genetic
Transcription Termination, Genetic
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Summary:The trigger loop (TL) forms a conserved element in the RNA polymerase active centre that functions in the elongation phase of transcription. Here, we show that the TL also functions in transcription initiation and termination. Using recombinant variants of RNA polymerase from Pyrococcus furiosus and a reconstituted transcription system, we demonstrate that the TL is essential for initial RNA synthesis until a complete DNA-RNA hybrid is formed. The archaeal TL is further important for transcription fidelity during nucleotide incorporation, but not for RNA cleavage during proofreading. A conserved glutamine residue in the TL binds the 2'-OH group of the nucleoside triphosphate (NTP) to discriminate NTPs from dNTPs. The TL also prevents aberrant transcription termination at non-terminator sites.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gkt433