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Structure of 5S rRNA within the Escherichia coli ribosome: Iodine-induced cleavage patterns of phosphorothioate derivatives

Published online by Cambridge University Press:  01 September 1998

OLGA V. SHPANCHENKO ,
OLGA A. DONTSOVA ,
ALEXEY A. BOGDANOV  and
KNUD H. NIERHAUS
OLGA V. SHPANCHENKO
Affiliation:
Department of Chemistry, Moscow State University, Moscow 119899, Russia Max-Planck-Institut für Molekulare Genetik, AG Ribosomen, Ihnestr. 73, D-14195 Berlin, Germany
OLGA A. DONTSOVA
Affiliation:
Department of Chemistry, Moscow State University, Moscow 119899, Russia
ALEXEY A. BOGDANOV
Affiliation:
Department of Chemistry, Moscow State University, Moscow 119899, Russia
KNUD H. NIERHAUS
Affiliation:
Max-Planck-Institut für Molekulare Genetik, AG Ribosomen, Ihnestr. 73, D-14195 Berlin, Germany
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Abstract

The protection patterns of 5S rRNA in solution, within the ribosomal 50S subunit, 70S ribosomes, and functional complexes, were assessed with the phosphorothioate method. About 20% of the analyzed positions (G9–G107) showed strong assembly defects: A phosphorothioate at one of these positions significantly impaired the incorporation of 5S rRNA into 50S particles. The reverse has also been observed: A phosphorothioate is preferred over a phosphate residue in the assembly process at a few positions. The results further demonstrate that 5S rRNA undergoes conformational changes during the assembly in the central protuberance of the 50S subunit and upon association with the small ribosomal subunit forming a 70S ribosome. In striking contrast, when the 70S ribosomes are once formed, the contact pattern of the 5S rRNA is the same in various functional states such as initiation-like complexes and pre- and posttranslocational states.

Keywords

phosphorothioate methodprotein synthesisribosomal assemblyribosomal elongation cycle5S rRNA
Type
Research Article
Information
RNA , Volume 4 , Issue 9 , September 1998 , pp. 1154 - 1164
Copyright
© 1998 RNA Society

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Footnotes

Reprint requests to: Knud H. Nierhaus, MPI für Molekulare Genetik, AG Ribosomen, Ihnestrasse 73, D-14195 Berlin, Germany; e-mail: nierhaus_kh@mpimg-berlin-dahlem.mpg.de.