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Yeast plasmid segregation: investigation of host/plasmid interactions

酵母质粒分离：宿主/质粒相互作用的研究

基本信息

批准号：
155268-2011
负责人：
Dobson, Melanie
金额：
$ 2.91万
依托单位：
Dalhousie University
依托单位国家：
加拿大
项目类别：
Discovery Grants Program - Individual
财政年份：
2014
资助国家：
加拿大
起止时间：
2014-01-01 至 2015-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=545658
关键词：
Yeast plasmid segregation investigation host

项目摘要

The 2µm circle is a high copy-number plasmid stably maintained in the nucleus of most strains of the bakers yeast, Saccharomyces cerevisiae. The 2µm circle is a "selfish" DNA, borrowing host machinery for its replication and segregation to ensure it is seldom lost when the host cell divides, while conferring no advantage to the host. We have identified sumoylation (modification of target proteins with a small ubiquitin-related protein called SUMO) as a host function that regulates plasmid segregation. Biochemical, molecular genetics, and cell biology approaches will be used to elucidate the link between sumoylation and plasmid maintenance and to identify and investigate other host proteins borrowed by the plasmid that enable its inheritance. Fluorescence microscopy will be used to investigate the effect of absence of host proteins identified in our studies on equal partitioning of the multiple plasmid copies at cell division and on plasmid localization within the nucleus. Our studies will also help identify the cellular process to which the identified host proteins contribute and reveal new functions for others. This project will determine how the 2µm circle interacts with other components of the nucleus to ensure its efficient segregation. Understanding how sumoylation and other host processes are exploited by the yeast plasmid may reveal how other genetic elements, such as viruses are maintained and help determine the essential role sumoylation plays in faithful segregation of chromosomes during cell division.

2μm 圆是一种高拷贝数质粒，稳定维持在大多数面包酵母菌株（酿酒酵母）的细胞核中。 2μm的圆圈是一种“自私”的DNA，借用宿主机制进行复制和分离，以确保它在宿主细胞分裂时很少丢失，同时不给宿主带来任何优势。我们已经确定苏修饰（用一种称为 SUMO 的小型泛素相关蛋白修饰靶蛋白）作为调节质粒分离的宿主功能。生物化学、分子遗传学和细胞生物学方法将用于阐明苏酰化和质粒维持之间的联系，并识别和研究质粒借用的其他宿主蛋白，使其能够遗传。荧光显微镜将用于研究我们研究中鉴定的宿主蛋白的缺失对细胞分裂时多个质粒拷贝的均等分配以及质粒在细胞核内定位的影响。我们的研究还将有助于确定已识别的宿主蛋白所贡献的细胞过程，并揭示其他蛋白的新功能。该项目将确定 2μm 圆如何与细胞核的其他成分相互作用，以确保其有效分离。了解酵母质粒如何利用苏酰化和其他宿主过程可能会揭示其他遗传元件（例如病毒）是如何维持的，并有助于确定苏酰化在细胞分裂过程中染色体忠实分离中所起的重要作用。