RRC ID 35103
著者 Sasano Y, Yamagishi K, Tanikawa M, Nakazawa T, Sugiyama M, Kaneko Y, Harashima S.
タイトル Stabilization of mini-chromosome segregation during mitotic growth by overexpression of YCR041W and its application to chromosome engineering in Saccharomyces cerevisiae.
ジャーナル J Biosci Bioeng
Abstract Chromosome engineering enables large-scale genome manipulation and can be used as a novel technology for breeding of yeasts. PCR-mediated chromosome splitting (PCS) offers a powerful tool for chromosome engineering by enabling a yeast chromosome to be split at any desired site. By applying PCS, a huge variety of chromosome combinations can be created and the best strain under specific conditions can be selected-a technology that we have called genome reorganization. Once the optimal strain is obtained, chromosome constitutions need to be maintained stably; however, mini-chromosomes of less than 50 kb are at relatively high frequency lost during cultivation. To overcome this problem, in this study we screened for multicopy suppressors of the high loss of mini-chromosomes by using a multicopy genomic library of Saccharomyces cerevisiae. We identified a novel gene, YCR041W, that stabilizes mini-chromosomes. The translational product of YCR041W was suggested to play an important role in increasing stability for mini-chromosome maintenance, probably by decreasing the rate of loss during mitotic cell division. The stabilization of mini-chromosomes conferred by YCR041W overexpression was completely dependent on the silencing protein Sir4, suggesting that a process related to telomere function might be involved in mini-chromosome stabilization. Overexpression of YCR041W stabilized not only a yeast artificial chromosome vector, but also a mini-chromosome derived from a natural chromosome. Taking these results together, we propose that YCR041W overexpression can be used as a novel chromosome engineering tool for controlling mini-chromosome maintenance and loss.
巻・号 119(5)
ページ 526-31
公開日 2015-5-1
DOI 10.1016/j.jbiosc.2014.10.006
PII S1389-1723(14)00384-3
PMID 25454064
MeSH Cell Cycle Proteins / genetics Cell Cycle Proteins / metabolism* Chromosome Segregation* Chromosomes, Artificial, Yeast / genetics* Chromosomes, Artificial, Yeast / metabolism Chromosomes, Fungal / genetics* Chromosomes, Fungal / metabolism Genomic Library Mitosis / genetics* Polymerase Chain Reaction Saccharomyces cerevisiae / cytology* Saccharomyces cerevisiae / genetics* Saccharomyces cerevisiae / metabolism Saccharomyces cerevisiae Proteins / genetics Saccharomyces cerevisiae Proteins / metabolism* Silent Information Regulator Proteins, Saccharomyces cerevisiae / genetics Silent Information Regulator Proteins, Saccharomyces cerevisiae / metabolism
IF 2.366
引用数 0
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY FOOD SCIENCE & TECHNOLOGY
リソース情報
酵母 NA