Nucleic Acids Research, 1992, Vol. 20, No. 15 4039-4046
© 1992
GENOME STRUCTURE AND MAPPING |
Characterization of Arabidopisis thaliana telomeres isolated in yeast
Cold spring Harbor Laboratory PO Box 100, 1 Bungtown Road, Cold Spring Harbor, NY 11724-2212, USA
*To whom Correspondence should be addressed
Received April 6, 1992. Revised June 2, 1992.
In an effort to learn more about the genomic organization of chromosomal termini in plants we employed a functional complementation strategy to isolate Arabidopsis thaliana telomeres in the yeast, Saccharomyces cerevisiae. Eight yeast eplsomes carrying A. thaliana telomeric sequences were obtained. The plant sequences carried on two episomes, YpAtTI and YpAtT7, were characterized in detail. The telomeric origins of YpAtTI and YpAtT7 Insert DNAs were confirmed by demonstrating that corresponding genomic sequences are preferentially degraded during exonucleolytic digestion. The isolated telomeric restriction fragments contain G-rich repeat arrays characteristic of A. thaliana telomeres, as well as subterminal telomere-associated sequences (TASs). DNA sequence analysis revealed the presence of variant telomeric repeats at the centromere-proximal border of the terminal block of telomere repeats. The TAS flanking the telomeric G-rich repeat in YpAtT7 corresponds to a repetitive element present at other A. thaliana telomeres, while more proximal sequences are unique to one telomere. The YpAtTI TAS is unique in the Landsberg strain of A. thaliana from which the clone originated; however, the Landsberg TAS cross-hybridizes weakly to a second telomere in the strain Columbia. Restriction analysis with cytosine methylation-sensitive endonucleases indicated that both TASs are highly methylated in the genome.
+Present address: Department of Genetics, Harvad Medical School, 25 Shattuck Street, Boston, MA 02115, USA
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