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Rapid Isolation of Centromeres from Scheffersomyces stipitis
Mingfeng Cao1, Arun Somwarpet Seetharam1, Andrew Josef Severin1
1Department of Chemical and Biological Engineering, ‡NSF Engineering Research Center for Biorenewable Chemicals (CBiRC), §Genome Informatics Facility, Office of Biotechnology, ∥Interdepartmental Microbiology Program, and ⊥The Ames Laboratory, 4140 Biorenewables Research Laboratory, Iowa State University , Ames, Iowa 50011, United States.
Researchers identified all eight centromeres (CENs) in yeast Scheffersomyces stipitis using a novel library-based strategy. This method efficiently isolates CENs by analyzing GC3 percentages, aiding in understanding chromosome stability.
Area of Science:
- Molecular Biology
- Genetics
- Yeast Biology
Background:
- Centromeres (CENs) are crucial for chromosome segregation and extrachromosomal DNA stability in yeasts.
- Identifying CENs in nonconventional yeast species is challenging due to their epigenetic nature.
- Previous work identified a 500-bp CEN5 in Scheffersomyces stipitis, confirming its role in plasmid stability.
Purpose of the Study:
- To develop an efficient strategy for isolating all centromeres (CENs) from Scheffersomyces stipitis.
- To investigate the potential of GC3 percentages for discerning centromere locations in yeast.
Main Methods:
- Integrated in silico GC3 chromosome scanning with high-throughput functional screening.
- Employed a library-based strategy for efficient isolation of centromeric sequences.
- Analyzed the characteristics of identified centromere core sequences.
Main Results:
- Successfully isolated all eight Scheffersomyces stipitis centromeres (CENs) with significant sequence reduction (16,000-fold).
- Identified a conserved 125-bp centromere core sequence present on all chromosomes.
- Observed that these core sequences are located in unique GC3-valley regions, suggesting GC3 percentage as a potential marker for CENs.
Conclusions:
- The developed library-based strategy efficiently isolates centromeres (CENs) in yeast.
- Local GC3 percentages can potentially serve as a reliable indicator for identifying centromere (CEN) locations in certain yeast species.
- This finding advances the understanding of centromere organization and identification in nonconventional yeasts.
