Highly selective affinity labelling of RNA polymerase B (II) from wheat germ.
DNA-dependent RNA polymerase B (II) from wheat germ was modified by incubation with 4-[N-(beta-hydroxyethyl)-N-methyl]benzaldehyde esters of AMP, ADP or ATP, followed by reduction with NaBH4. Reaction of the modified enzyme with [alpha-32P]UTP in the presence of various DNA templates led to a highly selective affinity labelling of the subunit with Mr 140 000 by covalently linked ApU. Labelling was inhibited by 1 microgram/ml alpha-amanitin.
M A
MA
Grachev
G R
GR
Hartmann
T G
TG
Maximova
A A
AA
Mustaev
A R
AR
Schäffner
H
H
Sieber
E F
EF
Zaychikov
D000345:Affinity Labels / Q000378:metabolism*; D000546:Amanitins / Q000494:pharmacology; D010761:Phosphorus Radioisotopes; D012319:RNA Polymerase II / Q000378:metabolism*; D014908:Triticum; D014544:Uridine Triphosphate / Q000378:metabolism
D000345:Affinity Labels; D000546:Amanitins; D010761:Phosphorus Radioisotopes; D012319:RNA Polymerase II; D014544:Uridine Triphosphate