Nucleic Acids Research, Vol 24, Issue 20 3996-4002, Copyright © 1996 by Oxford University Press
HA Meyer, F Triana-Alonso, CM Spahn, T Twardowski, A Sobkiewicz and KH Nierhaus
Antisense DNAs complementary against various sequences of the alpha- sarcin
domain (C2646-G2674) of 23S rRNA from Escherichia coli were hybridized to
naked 23S rRNA as well as to 70S ribosomes. Saturation levels of up to 0.4
per 70S ribosome were found, the identical fraction was susceptible to the
attack of the RNase alpha-sarcin. The hybridization was specific as
demonstrated with RNase H digestion, sequencing the resulting fragments and
blockage of the action of alpha- sarcin. The RNase alpha-sarcin seems to
approach its cleavage site from the 3' half of the loop of the alpha-sarcin
domain. Hybridization is efficiently achieved at 37 degrees C and can
extend at least into the 3' strand of the stem of the alpha-sarcin domain.
However, the inhibition of alpha-sarcin activity is observed at 30 degrees
C but not at 37 degrees C. For a significant inhibition of poly(Phe)
synthesis the temperature had to be lowered to 25 degrees C. The results
imply that the alpha-sarcin domain changes its conformation during protein
synthesis and that the conformational changes may include a melting of the
stem of the alpha-sarcin domain.
ARTICLES
Effects of antisense DNA against the alpha-sarcin stem-loop structure of the ribosomal 23S rRNA
Max-Planck-Institut fur Molekulare Genetik, AG Ribosomen, Berlin, Germany.
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