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J. Biol. Chem., Vol. 283, Issue 6, 3173-3180, February 8, 2008

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Translational Efficiency of a Non-AUG Initiation Codon Is Significantly Affected by Its Sequence Context in Yeast^*

Shun-Jia Chen, Grace Lin, Kuang-Jung Chang, Lu-Shu Yeh, and Chien-Chia Wang¹

From the Department of Life Science, National Central University, Jung-li, Taiwan 32001 and the Department of Life Science, Tzu-Chi University, Hua-lien, Taiwan 97041

Previous studies have shown that translation of mrna for yeast glycyl-tRNA synthetase is alternatively initiated from UUG and a downstream AUG initiation codon. Evidence presented here shows that unlike an AUG initiation codon, efficiency of this non-AUG initiation codon is significantly affected by its sequence context, in particular the nucleotides at positions –3 to –1 relative to the initiation codon. A/A/R (R represents A Or G) and C/G/C appear to be the most and least favorable sequences at these positions, respectively. Mutation of the native context sequence –3 to –1 from AAA to CGC reduced translation initiation from the UUG codon up to 32-fold and resulted in loss of mitochondrial respiration. although an AUG initiation codon is, in general, unresponsive to context changes in yeast, an AAA (–3 to –1) to CGC mutation still reduced its initiating activity up to 8-fold under similar conditions. these results suggest that sequence context is more important for translation initiation in yeast than previously appreciated.

Received for publication, August 20, 2007 , and in revised form, December 7, 2007.

^* This work was supported by Grant NSC 95-2311-B-008-005 and Grant NSC 95-2311-B-008-006 (to C.-C. W.) from the National Science Council (Taiwan). 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. Tel.: 886-3-426-0840; Fax: 886-3-422-8482; E-mail: dukewang{at}cc.ncu.edu.tw.

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