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Insights into telomerase action from high-throughput sequencing of S. pombe telomeres

Henrietta W. Bennett, Na Liu, Yan Hu, Megan C. King
doi: https://doi.org/10.1101/047837
Henrietta W. Bennett
1Department of Cell Biology, Yale School of Medicine, 333 Cedar Street, New Haven, CT, 06520-8002, USA
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Na Liu
1Department of Cell Biology, Yale School of Medicine, 333 Cedar Street, New Haven, CT, 06520-8002, USA
2Yale Stem Cell Center, 10 Amistad Street, New Haven, CT, 06520-8005, USA
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Yan Hu
1Department of Cell Biology, Yale School of Medicine, 333 Cedar Street, New Haven, CT, 06520-8002, USA
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Megan C. King
1Department of Cell Biology, Yale School of Medicine, 333 Cedar Street, New Haven, CT, 06520-8002, USA
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  • For correspondence: megan.king@yale.edu
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Abstract

We have developed a high-throughput sequencing approach that enables us to determine terminal telomere sequences from tens of thousands of individual Schizosaccharomyces pombe telomeres. This method provides unprecedented coverage of telomeric sequence complexity in fission yeast. S. pombe telomeres are composed of modular degenerate repeats that can be explained by variation in usage of the TER1 RNA template during reverse transcription. Taking advantage of this deep sequencing approach, we find that “like” repeat modules are highly correlated within individual telomeres. Moreover, repeat module preference varies with telomere length, suggesting that existing repeats promote the incorporation of like repeats and/or that specific conformations of the telomerase holoenzyme efficiently and/or processively add repeats of like nature. After the loss of telomerase activity, this sequencing and analysis pipeline defines a population of telomeres with altered sequence content. This approach should be adaptable to study telomeric repeats in other organisms and can provide new insights into telomere sequence content at high density.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted April 09, 2016.
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Insights into telomerase action from high-throughput sequencing of S. pombe telomeres
Henrietta W. Bennett, Na Liu, Yan Hu, Megan C. King
bioRxiv 047837; doi: https://doi.org/10.1101/047837
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Insights into telomerase action from high-throughput sequencing of S. pombe telomeres
Henrietta W. Bennett, Na Liu, Yan Hu, Megan C. King
bioRxiv 047837; doi: https://doi.org/10.1101/047837

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