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Title: Mapping of T7 RNA polymerase active site with novel reagents--oligonucleotides with reactive dialdehyde groups
Authors: Tunitskaya, V L
Rusakova, E E
Memelova, L V
Kochetkov, S N ×
Van Aerschot, Arthur
Herdewijn, Piet
Efimtseva, E V
Ermolinsky, B S
Mikhailov, S N #
Issue Date: Jan-1999
Publisher: Elsevier science bv
Series Title: FEBS Letters vol:442 issue:1 pages:20-4
Abstract: Oligonucleotides of a novel type containing 2'-O-beta-ribofuranosyl-cytidine were synthesized and further oxidized to yield T7 consensus promoters with dialdehyde groups. Both types of oligonucleotides were tested as templates, inhibitors, and affinity reagents for T7 RNA polymerase and its mutants. All oligonucleotides tested retained high affinity towards the enzyme. Wild-type T7 RNA polymerase and most of the mutants did not react irreversibly with oxidized oligonucleotides. Affinity labeling was observed only with the promoter-containing dialdehyde group in position (+2) of the coding chain and one of the mutants tested, namely Y639K. These results allowed us to propose the close proximity of residue 639 and the initiation region of the promoter within initiation complex. We suggest the oligonucleotides so modified may be of general value for the study of protein-nucleic acid interactions.
URI: 
ISSN: 0014-5793
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Medicinal Chemistry (Rega Institute)
× corresponding author
# (joint) last author

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