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Tetrahymena micronuclear sequences that function as telomeres in yeast.

J Shampay1, E H Blackburn

  • 1Department of Molecular Biology, University of California-Berkeley 94720.

Nucleic Acids Research
|April 25, 1989
PubMed
Summary

Baker

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Telomeres protect chromosome ends from degradation and fusion.
  • The sequence and structure of telomeric repeats are critical for telomere function.
  • Investigating heterologous telomere utilization provides insights into conserved mechanisms.

Purpose of the Study:

  • To determine if Saccharomyces cerevisiae (yeast) can use DNA sequences from Tetrahymena thermophila as functional telomeres.
  • To identify specific Tetrahymena DNA sequences that confer telomere function in yeast.
  • To explore the ability of yeast to heal broken chromosome ends using internal telomere-like sequences.

Main Methods:

  • Cloning of germline DNA from Tetrahymena thermophila into a linear yeast plasmid designed to select for telomere function.
  • Assaying the ability of cloned Tetrahymena DNA fragments to stabilize the linear plasmid in yeast.
  • Analyzing the sequence and location of functional Tetrahymena DNA elements within the yeast plasmid.

Main Results:

  • Only the (C4A2)n repeat sequences from Tetrahymena functioned as telomeres in yeast.
  • These (C4A2)n repeats did not need to originate from Tetrahymena telomeres; internal chromosomal fragments also worked.
  • Yeast elongated the functional Tetrahymena (C4A2)n repeats with its own telomeric sequences, and stabilized linear plasmids.

Conclusions:

  • Saccharomyces cerevisiae can recognize and utilize specific heterologous DNA sequences, namely (C4A2)n repeats, as functional telomeres.
  • The yeast system demonstrates broken chromosome healing by utilizing internally located telomere-like sequences.
  • This study highlights the importance of the telomeric repeat structure for in vivo recognition and function.

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