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Mutational analysis of the donor substrate binding site of the ribosomal peptidyltransferase center

Published online by Cambridge University Press:  01 February 1998

URMAS SAARMA ,
CHRISTIAN M.T. SPAHN ,
KNUD H. NIERHAUS  and
JAANUS REMME
URMAS SAARMA
Affiliation:
Institute of Molecular and Cell Biology, Tartu University, Riia 23, EE2400 Tartu Estonia
CHRISTIAN M.T. SPAHN
Affiliation:
Max-Planck-Institute für Molekulare Genetik, AG Ribosomen, Ihnestr. 73, D-14195 Berlin, Germany
KNUD H. NIERHAUS
Affiliation:
Max-Planck-Institute für Molekulare Genetik, AG Ribosomen, Ihnestr. 73, D-14195 Berlin, Germany
JAANUS REMME
Affiliation:
Institute of Molecular and Cell Biology, Tartu University, Riia 23, EE2400 Tartu Estonia
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Abstract

Previous experiments have shown that the top of helix 90 of 23S rRNA is highly important for the ribosomal peptidyltransferase activity and might be part of the donor (P) site. Developing on these studies, mutations in the 23S rRNA at the highly conserved positions G2505, G2582, and G2583 were investigated. None of the mutations affected assembly, subunit association, or the capacity of tRNA binding to A and P sites. A “selective transpeptidation assay” revealed that the mutations specifically impaired peptide bond formation. Results with a modified “fragment” assay using the minimal donor substrate pA-fMet are consistent with a model where the nucleotides ΨGG2582 form a binding pocket for C75 of the tRNA.

Keywords

Escherichia coli23S rRNA mutantsCMP stimulationthiostrepton
Type
Research Article
Information
RNA , Volume 4 , Issue 2 , February 1998 , pp. 189 - 194
Copyright
© 1998 RNA Society

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