[Study of the structure of Escherichia coli RNA polymerase and its complex with the lacUV5-promotor using protein-protein and DNA-protein crosslinks, formed by formaldehyde].
The protein-protein and DNA-protein crosslinks produced by formaldehyde were used to investigate the intersubunit and subunit-DNA interactions for free RNA polymerase and for an open complex of RNA polymerase with the lacUV5 promoter. In both cases the contacts between beta,beta' and beta', sigma subunits were observed, while there were no contacts between beta and sigma subunits. Only one of beta or beta' subunits and a sigma subunit crosslink to promoter DNA. We have chosen the conditions for fixing the RNA polymerase-DNA complexes on different stages of transcription initiation. The possibility to use limited fixation with low concentrations of formaldehyde to study specific DNA-protein interactions was shown.
Brodolin
K L
KL
Studitskiĭ
V M
VM
Mirzabekov
A D
AD
D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins / Q000737:chemistry*; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology*; D005557:Formaldehyde / Q000737:chemistry*; D011401:Promoter Regions, Genetic
D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D005557:Formaldehyde; D012321:DNA-Directed RNA Polymerases
Issledovanie struktury RNK-polimerazy Escherichia coli i ee komplekxa s lacUV5-promotorom s pomoshch'iu belok-belkovykh i DNK-belkovykh sshivok, obrazuemykh formal'degidom.