Translocation of sigma(70) with RNA polymerase during transcription: fluorescence resonance energy transfer assay for movement relative to DNA.
Using fluorescence resonance energy transfer, we show that, in the majority of transcription complexes, sigma(70) is not released from RNA polymerase upon transition from initiation to elongation, but, instead, remains associated with RNA polymerase and translocates with RNA polymerase. The results argue against the presumption that there are necessary subunit-composition differences, and corresponding necessary mechanistic differences, in initiation and elongation. The methods of this report should be generalizable to monitor movement of any molecule relative to any nucleic acid.
Mukhopadhyay
J
J
Howard Hughes Medical Institute, Waksman Institute, Department of Chemistry, Rutgers University, Piscataway, NJ 08854, USA.
Kapanidis
A N
AN
Mekler
V
V
Kortkhonjia
E
E
Ebright
Y W
YW
Ebright
R H
RH
D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D021381:Protein Transport; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014158:Transcription, Genetic
D012808:Sigma Factor; D004247:DNA; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases