A recombinant RNA polymerase II-like enzyme capable of promoter-specific transcription.
RNA polymerases (RNAPs) are core components of the cellular transcriptional machinery. Progress with functional studies of eukaryotic RNAPs has been delayed by the fact that it has not yet been possible to assemble active enzymes from individual subunits. Archaeal RNAPs are directly comparable to eukaryotic RNAPII in terms of primary sequence homology and quaternary structure. Here we report the successful in vitro assembly of a recombinant archaeal RNAP from purified subunits. The recombinant enzyme displays full activity in transcription assays and is capable, in the presence of two other basal factors, of promoter-specific transcription. The assembly of mutant enzymes yielded several unexpected insights into the structural and functional contributions of various subunits toward overall RNAP activity.
Werner
Finn
F
Department of Biological Sciences, Imperial College of Science, Technology and Medicine, London, United Kingdom.
Weinzierl
Robert O J
RO
D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001665:Binding Sites; D017017:Methanococcus / Q000201:enzymology; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic; D021122:Protein Subunits / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012319:RNA Polymerase II / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D014158:Transcription, Genetic
D019843:Archaeal Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D012319:RNA Polymerase II