title: T7 RNA polymerase functions in vitro without clustering.
abstract: Many nucleic acid polymerases function in clusters known as factories. We investigate whether the RNA polymerase (RNAP) of phage T7 also clusters when active. Using 'pulldowns' and fluorescence correlation spectroscopy we find that elongation complexes do not interact in vitro with a K(d)<1 µM. Chromosome conformation capture also reveals that genes located 100 kb apart on the E. coli chromosome do not associate more frequently when transcribed by T7 RNAP. We conclude that if clustering does occur in vivo, it must be driven by weak interactions, or mediated by a phage-encoded protein.
authors: Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom.
authors: Kieran
authors: K
authors: Finan
authors: Joseph P
authors: JP
authors: Torella
authors: Achillefs N
authors: AN
authors: Kapanidis
authors: Peter R
authors: PR
authors: Cook
mesh_terms: D017123:Bacteriophage T7 / Q000737:chemistry* / Q000378:metabolism; D002876:Chromosomes, Bacterial / Q000737:chemistry* / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases