Real time monitoring of the interaction of T7 RNA polymerase with azobenzene-tethered T7 promoter by biosensor.
We have already reported that transcription reaction by phage T7 or SP6 RNA polymerase (RNAP) can be reversibly photo-regulated with azobenzene-tethered promoter. Transcription reaction proceeded faster by UV irradiation than visible light irradiation. In the present study, binding of T7 RNAP to its azobenzene-tethered promoter was directly monitored by use of affinity biosensor. When azobenzene-tethered T7 promoter was immobilized on the sensor surface, response based on the binding of RNAP increased by UV light irradiation rather than by visible light irradiation. Thus, photo-regulation of transcription reaction by azobenzene-tethered promoter was attributed to the change of binding property of RNAP to the promoter by trans-cis isomerization of azobenzene.
Liu
Mingzhe
M
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Asanuma
Hiroyuki
H
Komiyama
Makoto
M
D001391:Azo Compounds / Q000737:chemistry / Q000378:metabolism*; D017123:Bacteriophage T7 / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D004279:DNA, Viral / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding / Q000528:radiation effects; D014466:Ultraviolet Rays; D014764:Viral Proteins / Q000378:metabolism*
D001391:Azo Compounds; D004279:DNA, Viral; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; C009850:azobenzene