PMID: 7537144Jan 1, 1995Paper

Circular rDNA replicons persist in Tetrahymena thermophila transformants synthesizing GGGGTC telomeric repeats

The Journal of Eukaryotic Microbiology
D P Romero, E H Blackburn

Abstract

Site-directed mutagenesis of the telomerase RNA from Tetrahymena thermophila was used previously to demonstrate the templating function of a sequence within this RNA; this sequence specifies the sequence of telomeric DNA in vivo. The possible functional importance of a phylogenetically conserved nucleotide outside the telomerase RNA template region was investigated by a similar experimental approach. The telomerase RNA gene was altered by site-directed mutagenesis, cloned in a circular selectable transformation vector consisting of an rRNA gene carrying a selectable drug resistance marker, and introduced into macronuclei of vegetatively dividing Tetrahymena thermophila by microinjection. Changing an invariant A to U at position 16 of the telomerase RNA (A16U) had no effect detectable by phenotype on telomerase function in vivo. However these experiments showed that a telomerase template alteration that dictates the synthesis of the mutant telomeric DNA sequence GGGGTC leads to a profound change in the population of rDNA replicons. The addition of GGGGTC mutant repeats leads to selective pressure for the loss of high copy linear rDNA, and the rRNA genes are maintained in the form of the circular rDNA replicons introduced during ...Continue Reading

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Citations

Aug 30, 2001·Current Biology : CB·S W ChanE H Blackburn
Jul 17, 1998·Journal of Molecular Biology·P Cohen, E H Blackburn
Apr 20, 1999·The Journal of Cell Biology·C D Smith, E H Blackburn
Jan 1, 1996·Molecular and Cellular Biology·D Gilley, E H Blackburn
Sep 15, 1995·Genes & Development·D GilleyE H Blackburn

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