title: Fluorescent methods to study transcription initiation and transition into elongation.
abstract: The DNA-dependent RNA polymerases induce specific conformational changes in the promoter DNA during transcription initiation. Fluorescence spectroscopy sensitively monitors these DNA conformational changes in real time and at equilibrium providing powerful ways to estimate interactions in transcriptional complexes and to assess how transcription is regulated by the promoter DNA sequence, transcription factors, and small ligands. Ensemble fluorescence methods described here probe the individual steps of promoter binding, bending, opening, and transition into the elongation using T7 phage and mitochondrial transcriptional systems as examples.
authors.affiliation: Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, Piscataway, NJ, 08854, USA.
authors.forename: Aishwarya P
authors.initials: AP
authors.lastname: Deshpande
authors.forename: Shemaila
authors.initials: S
authors.lastname: Sultana
authors.forename: Smita S
authors.initials: SS
authors.lastname: Patel
mesh_terms: D000818:Animals; D001665:Binding Sites; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D015336:Molecular Probe Techniques; D009690:Nucleic Acid Conformation; D061848:Optical Imaging / Q000379:methods*; D011401:Promoter Regions, Genetic; D013050:Spectrometry, Fluorescence; D061805:Transcription Elongation, Genetic; D061785:Transcription Initiation, Genetic; D014158:Transcription, Genetic
chemical_list: D005456:Fluorescent Dyes; D004247:DNA; D012321:DNA-Directed RNA Polymerases
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