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Nucleic Acids Research 2004 32(17):5206-5213; doi:10.1093/nar/gkh858
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Published online 30 September 2004

Nucleic Acids Research, Vol. 32 No. 17 © Oxford University Press 2004; all rights reserved

The Saccharomyces cerevisiae TRT2 tRNAThr gene upstream of STE6 is a barrier to repression in MAT{alpha} cells and exerts a potential tRNA position effect in MATa cells

Tiffany A. Simms, Elsy C. Miller, Nicolas P. Buisson, Nithya Jambunathan and David Donze*

Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA

* To whom correspondence should be addressed. Tel: +1 225 578 7391; Fax: +1 225 578 2597; Email: ddonze{at}lsu.edu

Received May 26, 2004; Revised and Accepted September 10, 2004

A growing body of evidence suggests that genes transcribed by RNA polymerase III exhibit multiple functions within a chromosome. While the predominant function of these genes is the synthesis of RNA molecules, certain RNA polymerase III genes also function as genomic landmarks. Transfer RNA genes are known to exhibit extra-transcriptional activities such as directing Ty element integration, pausing of replication forks, overriding nucleosome positioning sequences, repressing neighboring genes (tRNA position effect), and acting as a barrier to the spread of repressive chromatin. This study was designed to identify other tRNA loci that may act as barriers to chromatin-mediated repression, and focused on TRT2, a tRNAThr adjacent to the STE6 {alpha}2 operator. We show that TRT2 acts as a barrier to repression, protecting the upstream CBT1 gene from the influence of the STE6 {alpha}2 operator in MAT{alpha} cells. Interestingly, deletion of TRT2 results in an increase in CBT1 mRNA levels in MATa cells, indicating a potential tRNA position effect. The transcription of TRT2 itself is unaffected by the presence of the {alpha}2 operator, suggesting a hierarchy that favors assembly of the RNA polymerase III complex versus assembly of adjacent {alpha}2 operator-mediated repressed chromatin structures. This proposed hierarchy could explain how tRNA genes function as barriers to the propagation of repressive chromatin.


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