Open Access

Progressive rearrangement of telomeric sequences added to both the ITR ends of the yeast linear pGKL plasmid

  • Norio Gunge1Email author,
  • Hideki Takata2,
  • Akira Matsuura2 and
  • Kohsai Fukuda1
Biological Procedures Online5:51029

https://doi.org/10.1251/bpo44

Received: 2 December 2002

Accepted: 15 January 2003

Abstract

Relocation into the nucleus of the yeast cytoplasmic linear plasmids was studied using a monitor plasmid pCLU1. InSaccharomyces cerevisiae, the nuclearly-relocated pCLU1 replicated in a linear form (termed pTLU-type plasmid) which carried the host telomeric repeats TG1–3 of 300–350 bp at both ends. The telomere sequences mainly consisted of a major motif TGTGTGGGTGTGG which was complementary to part of the RNA template of yeast telomerase and were directly added to the very end of the pCLU1-terminal element ITR (inverted terminal repeat), suggesting that the ITR end played a role as a substrate of telomerase. The telomere sequences varied among isolated pTLU-type plasmids, but the TG1–3 organization was symmetrically identical on both ends of any one plasmid. During cell growth under non-selective condition, the telomeric repeat sequences were progressively rearranged on one side, but not on the opposite side of pTLU plasmid ends. This indicates that the mode of telomeric DNA replication or repair differed between both ends. Clonal analysis showed that the intense rearrangement of telomeric DNA was closely associated with extreme instability of pTLU plasmids.

Indexing terms

linear pGKL plasmidcytoplasmic linear plasmidtelomere de novo additiontelomerase Kluyveromyces lactis Saccharomyces cerevisiae

Notes

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