RRC ID 47255
Author Bennett HW, Liu N, Hu Y, King MC.
Title TeloPCR-seq: a high-throughput sequencing approach for telomeres.
Journal FEBS Lett
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 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.
Volume 590(23)
Pages 4159-4170
Published 2016-12-1
DOI 10.1002/1873-3468.12444
PMID 27714790
PMC PMC5561429
MeSH Base Sequence High-Throughput Nucleotide Sequencing / methods* Polymerase Chain Reaction Repetitive Sequences, Nucleic Acid / genetics Schizosaccharomyces / enzymology Schizosaccharomyces / genetics Telomerase / metabolism Telomere / genetics*
IF 3.057
Times Cited 2
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY BIOPHYSICS CELL BIOLOGY
Resource
Yeast wild-type S. pombe strain ?