Journal
FEBS LETTERS
Volume 590, Issue 23, Pages 4159-4170Publisher
WILEY
DOI: 10.1002/1873-3468.12444
Keywords
high-throughput sequencing; telomerase; telomere
Funding
- Yale Dean's Research Fellowship
- NSF [DBI-1156585]
- Raymond and Beverly Sackler Institute for Biological, Physical, and Engineering Sciences
- Yale Science, Technology and Research Scholars Program (STARS II)
- G. Harold and Leila Y. Mathers Charitable Foundation
- Searle Scholars Program
- New Innovator Award (National Institutes of Health, Office of the Director) [DP2OD008429]
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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 will be adaptable to study telomeric repeats in other organisms and also to interrogate repetitive sequences throughout the genome that are inaccessible to other sequencing methods.
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