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Molecular and Cellular Biology, June 2005, p. 4752-4766, Vol. 25, No. 11
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.11.4752-4766.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Identification of a Conserved RNA Motif Essential for She2p Recognition and mRNA Localization to the Yeast Bud{dagger}

Catherine Olivier,1 Guillaume Poirier,1 Patrick Gendron,2 Anita Boisgontier,2 François Major,2 and Pascal Chartrand1*

Département de Biochimie,1 Département d'Informatique et de Recherche Opérationnelle, Université de Montréal, Montréal, Canada2

Received 12 October 2004/ Returned for modification 15 November 2004/ Accepted 4 March 2005

In Saccharomyces cerevisiae, over twenty mRNAs localize to the bud tip of daughter cells, playing roles in processes as different as mating type switching and plasma membrane targeting. The localization of these transcripts depends on interactions between a cis-acting localization element(s) or zipcodes and the RNA-binding protein She2p. While previous studies identified four different localization elements in the bud-localized ASH1 mRNA, the main determinants for She2p recognition are still unknown. To investigate the RNA-binding specificity of She2p, we isolated She2p-binding RNAs by in vivo selection from libraries of partially randomized ASH1 localization elements. The RNAs isolated contained a similar loop-stem-loop structure with a highly conserved CGA triplet in one loop and a single conserved cytosine in the other loop. Mutating these conserved nucleotides or the stem separating them resulted in the loss of She2p binding and in the delocalization of a reporter mRNA. Using this information, we identified the same RNA motif in two other known bud-localized transcripts, suggesting that this motif is conserved among bud-localized mRNAs. These results show that mRNAs with zipcodes lacking primary sequence similarity can rely on a few conserved nucleotides properly oriented in their three-dimensional structure in order to be recognized by the same localization machinery.

* Corresponding author. Mailing address: Département de Biochimie, Université de Montréal, Montréal, QC H3C 3J7, Canada. Phone: (514) 343-5684. Fax: (514) 343-2210. E-mail: p.chartrand{at}umontreal.ca.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, June 2005, p. 4752-4766, Vol. 25, No. 11
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.11.4752-4766.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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