Static and kinetic site-specific protein-DNA photocrosslinking: analysis of bacterial transcription initiation complexes.
Static site-specific protein-DNA photocrosslinking permits identification of protein-DNA interactions within multiprotein-DNA complexes. Kinetic site-specific protein-DNA photocrosslinking - involving rapid-quench-flow mixing and pulsed-laser irradiation - permits elucidation of pathways and kinetics of formation of protein-DNA interactions within multiprotein-DNA complexes. We present detailed protocols for application of static and kinetic site-specific protein-DNA photocrosslinking to bacterial transcription initiation complexes.
Naryshkin
Nikolai
N
HHMI, Waksman Institute, Rutgers University, 190 Frelinghuysen Road, Piscataway, NJ 08854, USA.
Druzhinin
Sergei
S
Revyakin
Andrei
A
Kim
Younggyu
Y
Mekler
Vladimir
V
Ebright
Richard H
RH
D000818:Animals; D003432:Cross-Linking Reagents / Q000737:chemistry* / Q000528:radiation effects*; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D007700:Kinetics; D008027:Light; D008967:Molecular Biology / Q000379:methods*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics*
D003432:Cross-Linking Reagents; D011506:Proteins; D004247:DNA; D012321:DNA-Directed RNA Polymerases