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Molecular and Cellular Biology, May 2008, p. 3301-3312, Vol. 28, No. 10
0270-7306/08/$08.00+0     doi:10.1128/MCB.01542-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Eukaryotic Wobble Uridine Modifications Promote a Functionally Redundant Decoding System{triangledown}

Marcus J. O. Johansson,{dagger} Anders Esberg,{ddagger} Bo Huang, Glenn R. Björk, and Anders S. Byström*

Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden

Received 23 August 2007/ Returned for modification 23 October 2007/ Accepted 27 February 2008

The translational decoding properties of tRNAs are modulated by naturally occurring modifications of their nucleosides. Uridines located at the wobble position (nucleoside 34 [U34]) in eukaryotic cytoplasmic tRNAs often harbor a 5-methoxycarbonylmethyl (mcm5) or a 5-carbamoylmethyl (ncm5) side chain and sometimes an additional 2-thio (s2) or 2'-O-methyl group. Although a variety of models explaining the role of these modifications have been put forth, their in vivo functions have not been defined. In this study, we utilized recently characterized modification-deficient Saccharomyces cerevisiae cells to test the wobble rules in vivo. We show that mcm5 and ncm5 side chains promote decoding of G-ending codons and that concurrent mcm5 and s2 groups improve reading of both A- and G-ending codons. Moreover, the observation that the mcm5U34- and some ncm5U34-containing tRNAs efficiently read G-ending codons challenges the notion that eukaryotes do not use U-G wobbling.

* Corresponding author. Mailing address: Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden. Phone: 46 90 785 67 64. Fax: 46 90 77 26 30. E-mail: Anders.Bystrom{at}molbiol.umu.se

{triangledown} Published ahead of print on 10 March 2008.

{dagger} Present address: Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA 01655.

{ddagger} Present address: Department of Molecular Genetics and Microbiology, 3020 Duke University Medical Center, Durham, NC 27710.

Molecular and Cellular Biology, May 2008, p. 3301-3312, Vol. 28, No. 10
0270-7306/08/$08.00+0     doi:10.1128/MCB.01542-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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