title: Role of open complex instability in kinetic promoter selection by bacteriophage T7 RNA polymerase.
abstract: By measuring steady-state rates of dinucleotide synthesis on double-stranded (d.s.) and partially single-stranded (p.s.s.) promoters, and topological unwinding due to open complex formation on plasmids, we have obtained evidence that open complex formation in bacteriophage T7 RNA polymerase:promoter binary complexes is thermodynamically disfavored and that the rate of collapse of the open complex is competitive with the rate of transcription initiation. It is suggested that open complex instability is a kinetic mechanism that allows T7 RNA polymerase (RNAP) to achieve promoter specificity while still allowing for efficient promoter release. Open complex instability could also provide a mechanism for modulating the KM for the initiating NTPs so as to allow different promoters to respond differently to physiological changes in NTP concentration.
authors.affiliation: Department of Biochemistry, University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78212-7760, USA.
authors.forename: J
authors.initials: J
authors.lastname: Villemain
authors.forename: R
authors.initials: R
authors.lastname: Guajardo
authors.forename: R
authors.initials: R
authors.lastname: Sousa
mesh_terms: D017123:Bacteriophage T7 / Q000201:enzymology*; D004247:DNA / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D011401:Promoter Regions, Genetic; D013379:Substrate Specificity; D013816:Thermodynamics; D014158:Transcription, Genetic; D014764:Viral Proteins
chemical_list: D004277:DNA, Single-Stranded; D004279:DNA, Viral; D014764:Viral Proteins; D006160:Guanosine Triphosphate; D004247:DNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases