Telomere Sequences Attached to Nuclearly Migrated Yeast Linear Plasmid

• Published in: Plasmid 2000-03, Vol.43 (2), p.137-143
• Main Authors: Takata, Hideki, Fukuda, Kohsai, Meinhardt, Friedhelm, Gunge, Norio
• Format: Article
• Language: English
• Subjects: Base Sequence; Cell Nucleus - chemistry; Cell Nucleus - genetics; Consensus Sequence; cytoplasm; DNA, Fungal - genetics; DNA, Fungal - isolation & purification; DNA-Binding Proteins - deficiency; DNA-Binding Proteins - genetics; Molecular Sequence Data; Plasmids - chemistry; Plasmids - isolation & purification; Rad52 DNA Repair and Recombination Protein; RAD52 gene; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins; Sequence Alignment; Sequence Analysis, DNA; Telomere - chemistry; Telomere - genetics
• ISSN: 0147-619X; 1095-9890
• DOI: 10.1006/plas.1999.1454

The yeast linear plasmid pCLU1, derived from pGKL1, has terminal proteins (TPs) covalently attached at the 5′ ends of inverted terminal repeats (ITRs) and replicates in the cytoplasm, presumably using the TP as a primer for DNA synthesis. In Saccharomyces cerevisiae, under certain conditions, pCLU1 migrated into the nucleus and replicated in either linear or circular form. The linear-form plasmid lacked TPs; instead it carried host-telomere repeats at the ITR ends. The present study showed that (1) the added telomere was primarily composed of the repeated tracts of TGTGTGGGTGTGG, which was complementary to the RNA template of yeast telomerase, (2) the telomeric addition occurred at the very end of the ITRs, and (3) the sequence composition of the added telomeres was diverse among individual plasmids, but symmetrically identical at both ends of each plasmid. A similar mode of telomere addition was also observed in cells defective in the RAD52 gene.