Real time in vitro analysis of transcription by RNA polymerase on immobilized DNA fibres.
We have used surface plasmon resonance (SPR) to follow variations in the concentrations of binary complexes as RNA polymerase moves into a transcriptionally competent initiation complex with immobilized DNA fibres containing promoter sequences. The use of SPR to follow complex binding phenomena is described. We have also followed the changes in the mass of initiation complexes following addition of the nucleotide triphosphates prerequisite for transcription on the immobilized template. These signals are interpreted in terms of the escape of RNA polymerase into elongation mode and the subsequent synthesis of nascent RNA molecules.
Pemberton
I K
IK
Unité de Physicochimie des Macromolécules Biologiques (URA 1149 du CNRS), Institut Pasteur, 25 Rue du Dr Roux, 75724 Paris cédex 15, France.
Buckle
M
M
D004247:DNA / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D020869:Gene Expression Profiling; D011401:Promoter Regions, Genetic; D020349:Surface Plasmon Resonance / Q000379:methods*; D013698:Templates, Genetic; D013997:Time Factors; D014158:Transcription, Genetic
D004247:DNA; D012321:DNA-Directed RNA Polymerases