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Telomere sequences attached to nuclearly migrated yeast linear plasmid.

H Takata¹, K Fukuda, F Meinhardt

¹Kumamoto Institute of Technology, Ikeda 4-22-1, Kumamoto, 860-0082, Japan.

|February 25, 2000

Summary

The yeast linear plasmid pCLU1, when in the nucleus, loses its terminal proteins (TPs) and gains host telomere repeats at its ends. This telomere addition is guided by yeast telomerase and occurs symmetrically.

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Area of Science:

Molecular Biology
Yeast Genetics
Telomere Biology

Background:

The yeast linear plasmid pCLU1 replicates in the cytoplasm using terminal proteins (TPs) as primers.
Under certain conditions, pCLU1 can enter the nucleus and replicate as a linear or circular molecule.
Nuclear linear pCLU1 lacks TPs and instead acquires host telomere repeats at its inverted terminal repeats (ITRs).

Purpose of the Study:

To characterize the telomere addition process in nuclear linear pCLU1.
To determine the sequence and location of added telomeres.
To investigate the mechanism of telomere acquisition in relation to yeast telomerase and DNA repair pathways.

Main Methods:

Analysis of pCLU1 DNA sequences from Saccharomyces cerevisiae.

Related Experiment Videos

DNA sequencing to identify telomere repeat composition and attachment sites.

Comparison of telomere addition in wild-type and RAD52-deficient cells.

Main Results:

The added telomeres primarily consist of TGTGTGGGTGTGG repeats, complementary to yeast telomerase RNA.
Telomeric addition occurs precisely at the ends of the inverted terminal repeats (ITRs).
Telomere sequences are diverse among plasmids but symmetrically identical at both ends of each plasmid.
Similar telomere addition was observed in RAD52-deficient cells, suggesting a RAD52-independent mechanism.

Conclusions:

Nuclear linear pCLU1 acquires telomeres via a mechanism involving yeast telomerase.
Telomere addition occurs at the ITRs and is symmetrically established.
The process appears to be independent of the RAD52 gene, a key component of homologous recombination.