Nucleic Acids Research, 1989, Vol. 17, No. 8 3247-3260
© 1989
MOLECULAR BIOLOGY |
Tetrahymena micronuclear sequences that function as telomeres in yeast
Department of Molecular Biology, University of California-Berkeley Berkeley, CA 94720. USA
*To whom correspondence should be addressed
Received September 19, 1988. Revised March 8, 1989. Accepted March 8, 1989.
We explored the ability of S. cerevisiae to utilize heterologous DNA sequences as telomeres by cloning germline (micronuclear) DNA from Tetrahymena thermophila on a linear yeast plasmid that selects for telomere function. The only Tetrahymena sequences that functioned in this assay were (C4A2)n repeats. Moreover, these repeats did not have to be derived from Tetrahymena telomeres, although we show that micronuclear telomeres (like macronuclear telomeres) of Tetrahymena terminate in (C4A2)n repeats. Chromosome-internal restriction fragments carrying (C4A2)n repeats also stabilized linear plasmids and were elongated by yeast telomeric repeats. In one case, the C4A2 repeat tract was approximately 1.5 kb from the end of the genomic Tetrahymena DNA fragment that was cloned, but this 1.5 kb of DNA was missing from the linear plasmid. Thus, yeast can utilize internally located tracts of telomere-like sequences, after the distal DNA is removed. The data provide an example of broken chromosome healing, and underscore the importance of the telomeric repeat structure for recognition of functional telomeric DNA in vivo.
+Present address: HSE 1556, University of California-San Francisco, San Francisco, CA 94143-0554, USA
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