Nucleic Acids Research, 2001, Vol. 29, No. 3 710-715
© 2001 Oxford University Press
Mutagenesis of the peptidyltransferase center of 23S rRNA: the invariant U2449 is dispensable
J. W. Wilson Laboratory, Department of Molecular and Cellular Biology and Biochemistry, Brown University, Providence, RI 02912, USA and 1Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111, USA
U2449 is one of many invariant residues in the central loop of domain V of 23S rRNA, a region that constitutes part of the peptidyltransferase center of the ribosome. In Escherichia coli, this U is post-transcriptionally modified to dihydrouridine (D) and is the only D modification found in E.coli rRNAs. To analyze the role of this base and its modification in ribosomal function, all three base substitutions were constructed on a plasmid copy of the rrnB operon and assayed for their ability to support cell growth in a strain of E.coli lacking chromosomal rrn operons. Both purine substitution mutations were not viable. However, growth and antibiotic sensitivity of cells expressing only the mutant D2449C rRNA was indistinguishable from wild type. We conclude that while a pyrimidine is required at position 2449 for proper ribosomal function, the D modification is dispensable.
* To whom correspondence should be addressed. Tel: +1 401 863 3652; Fax: +1 401 863 1182; Email: michael_o'connor{at}brown.edu
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