[Highly selective labeling of the E. coli RNA-polymerase promoter complex with reactive derivatives of oligonucleotide primers of various specificity].
A technique of highly selective affinity labelling, which includes covalent modification of the enzyme-T7A2 promoter complex with reactive oligonucleotide derivatives and subsequent elongation of the attached oligonucleotide residue with a radioactive substrate was used to study the product-binding site of E. coli RNA polymerase. Different oligonucleotides complementary to the T7A2 promoter (with lengths ranging from 2 to 8 residues) containing 5'-terminal phosphorylating, alkylating or aldehyde groups were used for the labelling. The procedure resulted in labelling DNA and beta-, beta'- or sigma-subunits of the enzyme, which are therefore believed to contact with growing RNA in the course of initiation. Consideration of the labelling patterns as a functions of the oligonucleotide's length as well as of the structure and chemical specificity of the reactive groups led to a tentative topographic scheme of the RNA polymerase product-binding region.
Tsarev
I G
IG
Mustaev
A A
AA
Zaĭchikov
E F
EF
Alikina
T Iu
TIu
Ven'iaminova
A G
AG
Repkova
M N
MN
D000345:Affinity Labels / Q000138:chemical synthesis; D001483:Base Sequence; D001665:Binding Sites; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011401:Promoter Regions, Genetic; D013379:Substrate Specificity; D013698:Templates, Genetic
D000345:Affinity Labels; D015345:Oligonucleotide Probes; D012321:DNA-Directed RNA Polymerases
Vysokoselektivnoe mechenie kompleksa RNK-polimerazy E. coli s promotorom reaktsionnosposobnymi proizvodnymi oligonukleotidov- zatravok razlichnoĭ spetsifichnosti.