DNA plasmid, transmission in yeast is associated with specific sub-nuclear localisation during cell division
DNA plasmid, transmission in yeast is associated with specific sub-nuclear localisation during cell division
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DOI：
10.1006/cbir.2002.0867
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发表时间：
2002-01-01
影响因子：
3.9
通讯作者：
Murray, JAH
中科院分区：
文献类型：
--
作者：
Scott-Drew, S;Wong, CMVL;Murray, JAH
酵母中仅携带DNA复制起点（ARS）的环状质粒表现出母性遗传偏好（MIB），并且在分裂过程中从母细胞到子细胞的传递很差。已经描述了克服MIB的各种不同序列，包括着丝粒序列（CEN）、端粒相关重复序列、沉默序列和由内源性2 μ环质粒编码的需要顺式作用基因座STB和蛋白质Rep1和Rep2的特定系统。在每种情况下，母细胞和子细胞之间的DNA分离取决于DNA-蛋白质相互作用。使用质粒携带多个拷贝的乳糖阻遏物结合序列，我们本地化的DNA分子在酵母细胞核中使用绿色荧光蛋白（GFP）-乳糖阻遏物融合蛋白。我们比较了携带着丝粒序列的GFP定位质粒与基于2 μ环的质粒，所述质粒携带或缺乏分离所需的STB序列。我们发现，GFP本地化质粒携带完整的STB基因座与质粒蛋白Rep1和Rep2的离散染色质位点共定位。这些位点既不同于端粒，也不同于粘着蛋白结合位点。删除STB区域对质粒的稳定性至关重要，导致质粒与染色质结合的丧失，质粒向核周边的重新定位，以及与质粒结合的Rep I蛋白的减少。我们的结论是，特定的质粒定位可能是重要的克服MIB在酵母中。(C)2002爱思唯尔科技有限公司。保留所有权利。
Circular plasmids in yeast carrying only an origin Of DNA replication (ARS) exhibit maternal inheritance bias (MIB) and are poorly transmitted from mother to daughter cell during division. A variety of different sequences that overcome MIB have been described, including centromeric sequences (CEN), telomere-associated repeats, silencer sequences and a specific system encoded by the endogenous 2 mu circle plasmid requiring the cis-acting locus STB and the proteins Rep1 and Rep2. In each case, DNA segregation between mother and daughter cells is dependent on DNA-protein interactions. Using plasmids carrying multiple copies of a lac repressor binding sequence, we have localised DNA molecules in the yeast nucleus using a green fluorescent protein (GFP)-lac repressor fusion protein. We compared GFP localised plasmids carrying a centromere sequence with plasmids based on 2 mu circle carrying or lacking the STB sequences required for their segregation. We show that GFP localised plasmid carrying the complete STB locus co-localises with the plasmid proteins Rep1 and Rep2 to discrete chromatin sites. These sites are distinct from both the telomeres and from sites of cohesin binding. Deletion of the region of STB essential for the stability of the plasmid, leads to a loss of plasmid association with chromatin, relocalisation of plasmids towards the nuclear periphery, and a decrease in the Rep I protein associated with the plasmid. We conclude that specific plasmid localisation is likely to be important in the overcoming of MIB in yeast. (C) 2002 Elsevier Science Ltd. All rights reserved.