Prajapati Hemant Kumar, Rizvi Syed Meraj Azhar, Rathore Ishan, Ghosh Santanu K
Department of Biosciences and Bioengineering, Indian Institute of Technology, Bombay, Powai, Mumbai, 400076, India.
Mol Microbiol. 2017 Mar;103(6):1046-1064. doi: 10.1111/mmi.13608. Epub 2017 Jan 15.
The 2 μ plasmid of budding yeast shows high mitotic stability similar to that of chromosomes by using its self-encoded systems, namely partitioning and amplification. The partitioning system consists of the plasmid-borne proteins Rep1, Rep2 and a cis-acting locus STB that, along with several host factors, ensures efficient segregation of the plasmid. The plasmids show high stability as they presumably co-segregate with chromosomes through utilization of various host factors. To acquire these host factors, the plasmids are thought to localize to a certain sub-nuclear locale probably assisted by the motor protein, Kip1 and microtubules. Here, we show that the microtubule-associated proteins Bik1 and Bim1 are also important host factors in this process, perhaps by acting as an adapter between the plasmid and the motor and thus helping to anchor the plasmid to microtubules. Abrogation of Kip1 recruitment at STB in the absence of Bik1 argues for its function at STB upstream of Kip1. Consistent with this, both Bik1 and Bim1 associate with plasmids without any assistance from the Rep proteins. As observed earlier with other host factors, lack of Bik1 or Bim1 also causes a cohesion defect between sister plasmids leading to plasmid missegregation.
出芽酵母的2μ质粒通过其自身编码的系统,即分配和扩增,表现出与染色体相似的高有丝分裂稳定性。分配系统由质粒携带的蛋白质Rep1、Rep2和一个顺式作用位点STB组成,该位点与几个宿主因子一起确保质粒的有效分离。这些质粒表现出高稳定性,因为它们可能通过利用各种宿主因子与染色体共同分离。为了获得这些宿主因子,这些质粒被认为可能在驱动蛋白Kip1和微管的协助下定位到特定的核内亚区域。在这里,我们表明微管相关蛋白Bik1和Bim1也是这一过程中的重要宿主因子,可能是通过充当质粒与驱动蛋白之间的衔接子,从而帮助将质粒锚定到微管上。在没有Bik1的情况下,STB处Kip1募集的缺失证明了它在Kip1上游的STB处发挥作用。与此一致的是,Bik1和Bim1在没有Rep蛋白的任何协助下都与质粒相关联。正如早期对其他宿主因子的观察一样,缺乏Bik1或Bim1也会导致姐妹质粒之间的黏连缺陷,从而导致质粒错误分离。