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J. Biol. Chem., Vol. 281, Issue 40, 29850-29862, October 6, 2006

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Mutations in the Intersubunit Bridge Regions of 23 S rRNA^*

Aivar Liiv and Michael O'Connor¹

From the Estonian Biocentre, Tartu University, Tartu 51010, Estonia and the School of Biological Sciences, University of Missouri, Kansas City, Missouri 64110

The large and small subunits of the ribosome are joined by a series of bridges that are conserved among mitochondrial, bacterial, and eukaryal ribosomes. In addition to joining the subunits together at the initiation of protein synthesis, a variety of other roles have been proposed for these bridges. These roles include transmission of signals between the functional centers of the two subunits, modulation of tRNA-ribosome and factor-ribosome interactions, and mediation of the relative movement of large and small ribosomal subunits during translocation. The majority of the bridges involve RNA-RNA interactions, and to gain insight into their function, we constructed mutations in the 23 S rRNA regions involved in forming 7 of the 12 intersubunit bridges in the Escherichia coli ribosome. The majority of the mutants were viable in strains expressing mutant rRNA exclusively but had distinct growth phenotypes, particularly at 30 °C, and the mutant ribosomes promoted a variety of miscoding errors. Analysis of subunit association activities both in vitro and in vivo indicated that, with the exception of the bridge B5 mutants, at least one mutation at each bridge site affected 70 S ribosome formation. These results confirm the structural data linking bridges with subunit-subunit interactions and, together with the effects on decoding fidelity, indicate that intersubunit bridges function at multiple stages of protein synthesis.

Received for publication, March 30, 2006 , and in revised form, August 4, 2006.

^* This work was supported by Grant MCB 0343942 from the National Science Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: School of Biological Sciences, University of Missouri, 5007 Rockhill Rd., Kansas City, MO 64110. Tel.: 816-235-6372; Fax: 816-235-5595; E-mail: oconnormi{at}umkc.edu.

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