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Vol. 12, No. 20, pp. 3286-3300, October 15, 1998
Departments of Microbiology and Immunology and Biochemistry and
Biophysics, University of California, San Francisco, San Francisco,
California 94143-0414 USA
The reverse transcriptase telomerase is a ribonucleoprotein complex
that adds telomeric repeats to chromosome ends, using a sequence within
its endogenous RNA component as a template. Although templating domains
of telomerase RNA have been studied in detail, little is known about
the roles of the remaining residues, particularly in yeast. We examined
the functions of nontemplate telomerase residues in the telomerase RNA
of budding yeast Kluyveromyces lactis. Although approximately
half of the RNA residues were dispensable for function, four specific
regions were essential for telomerase action in vivo. We analyzed the
effects of mutating these regions on in vivo function, in vitro
telomerase activity, and telomerase RNP assembly. Deletion of two
regions resulted in synthesis of stable RNAs that appeared unable to
assemble into a stable RNP. Mutating a region near the 5' end of
the RNA allowed RNP assembly but abolished enzymatic activity.
Mutations in another specific small region of the RNA led to an
inactive telomerase RNP with an altered RNA conformation.
[Key Words: Telomerase; RNA; ribonucleoprotein; telomere; yeast; K. lactis]
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