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Molecular and Cellular Biology, April 2002, p. 2366-2374, Vol. 22, No. 7
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.7.2366-2374.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Essential Regions of Saccharomyces cerevisiae Telomerase RNA: Separate Elements for Est1p and Est2p Interaction

April J. Livengood,1 Arthur J. Zaug,2,3 and Thomas R. Cech1,2,3*

Department of MCD Biology,1 Department of Chemistry and Biochemistry,2 Howard Hughes Medical Institute, University of Colorado, Boulder, Colorado 80309-02153

Received 20 September 2001/ Returned for modification 1 November 2001/ Accepted 3 January 2002

The Saccharomyces cerevisiae telomerase RNA subunit is encoded by the TLC1 gene. A selection for viable alleles of TLC1 RNA from a large library of random deletion alleles revealed that less than half (~0.5 kb of the ~1.3-kb RNA) is required for telomerase function in vivo. The main essential region (430 nucleotides), which contains the template for telomeric DNA synthesis, was required for coimmunoprecipitation with Est1p and Est2p. Furthermore, the subregion required for interaction with Est1p, the telomerase recruitment subunit, differed from those required for interaction with Est2p, the reverse transcriptase subunit. Two regions of the RNA distant from the template in the nucleotide sequence were required for Est2p binding, but the template itself was not. Having the RNA secured to the protein away from the template is proposed to facilitate the translocation of the RNA template through the active site. More generally, our results support a role for the telomerase RNA serving as a scaffold for binding key protein subunits.

* Corresponding author. Mailing address: Howard Hughes Medical Institute, 4000 Jones Bridge Rd., Chevy Chase, MD 20815. Phone: (301) 215-8550. Fax: (301) 215-8558. E-mail: CECHT{at}hhmi.org.

Molecular and Cellular Biology, April 2002, p. 2366-2374, Vol. 22, No. 7
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.7.2366-2374.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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