Nucleic Acids Research, Vol 26, Issue 2 549-553, Copyright © 1998 by Oxford University Press
RA Poot, SH van den Worm, CWA Pleij and J van Duin
Helix 2 of the central pseudoknot structure in Escherichia coli 16S rRNA is
formed by a long-distance interaction between nt 17-19 and 918- 916,
resulting in three base pairs: U17-A918, C18-G917and A19-U916. Previous
work has shown that disruption of the central base pair abolishes ribosomal
activity. We have mutated the first and last base pairs and tested the
mutants for their translational activity in vivo , using a specialized
ribosome system. Mutations that disrupt Watson- Crick base pairing result
in strongly impaired translational activity. An exception is the mutation
U916-->G, creating an A.G pair, which shows almost no decrease in
activity. Mutations that maintain base complementarity have little or no
impact on translational efficiency. Some of the introduced base pair
substitutions substantially alter the stability of helix 2, but this does
not influence ribosome functioning, neither at 42 nor at 28 degrees C.
Therefore, our results do not support models in which the pseudoknot is
periodically disrupted. Rather, the central pseudoknot structure is
suggested to function as a permanent structural element necessary for
proper organization in the center of the 30S subunit.
ARTICLES
Base complementarity in helix 2 of the central pseudoknot in 16S rRNA is essential for ribosome functioning
Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, PO Box 9502, 2300 RA Leiden, The Netherlands.
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