[Point contacts of T7 RNA polymerase in the promotor complex, as determined with phosphate-activated oligonucleotide derivatives].
The contacts between phosphate groups of promoter DNA an Lys or His of T7 RNA polmerase (Pol) in the Pol-promoter complex were studied with single- and double- stranded oligonucleotides, which corresponded to the T7 promoter consensus and contained activated phosphate groups at position +1, +2, or -14 relative to the transcription start. To obtain reactive groups, terminal phosphates were modified with N-oxybenzotriazole (HOBT), and internucleotide phosphates were repalced with a trisubstituted pyrophosphate (TSP). The resulting derivatives produced covalent complexes with T7 Pol. Covalent bonding involved His in the case of TSP at position +1 or HOBT at position +1 or -14, and Lys in the case of TSB at position -14.
Filippova
S E
SE
Belozerskii Institute of Physico-Chemical Biology, Moscow Sate University, Moscow, 119899 Russia.
Ivanovskaia
M G
MG
Romanova
E A
EA
Tunitskaia
V L
VL
Kochetkov
S N
SN
D001665:Binding Sites; D001671:Biochemistry / Q000379:methods; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011756:Diphosphates / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D010710:Phosphates / Q000737:chemistry*; D011401:Promoter Regions, Genetic; D014764:Viral Proteins
D011756:Diphosphates; D010710:Phosphates; D014764:Viral Proteins; D006639:Histidine; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D008239:Lysine