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Rapid Isolation of Centromeres from Scheffersomyces stipitis

TitleRapid Isolation of Centromeres from Scheffersomyces stipitis
Publication TypeJournal Article
Year of Publication2017
AuthorsCao, MF, Seetharam, AS, Severin, AJ, Shao, ZY
JournalAcs Synthetic Biology
Volume6
Pagination2028-2034
Date Published11
Type of ArticleArticle
ISBN Number2161-5063
Accession NumberWOS:000416204100007
Keywordsbudding yeast, candida-albicans, centromeres, chromosomes, construction, dna assembler, episomal plasmids, evolution, GC(3) chromosome scanning epigeneticity, genome, Molecular Biology, nonconventional yeasts, nucleosome, pichia-stipitis, Scheffersomyces stipitis, sequences
Abstract

Centromeres (CENs) are the chromosomal regions promoting kinetochore formation for faithful chromosome segregation. In yeasts, CENs have been recognized as the essential elements for extra-chromosomal DNA stabilization. However, the epigeneticity of CENs makes their localization on individual chromosomes very challenging, especially in many not well-studied nonconventional yeast species. Previously, we applied a stepwise method to identify a 500-bp CENS from Scheffersomyces stipitis chromosome and experimentally confirmed its critical role on improving plasmid stability. Here we report a library-based strategy that integrates in silico GC(3) chromosome scanning and high-throughput functional screening, which enabled the isolation of all eight S. stipitis centromeres with a 16 000-fold reduction in sequence very efficiently. Further identification of a 125-bp CEN core sequence that appears multiple times on each chromosome but all in the unique signature GC(3)-valley indicates that CEN location might be accurately discerned by their local GC(3) percentages in a subgroup of yeasts.

DOI10.1021/acssynbio.7b00166
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