Ortiz J, Stemmann O, Rank S, Lechner J
Institut für Biochemie, Genetik und Mikrobiologie, Universität Regensburg, 93040 Regensburg, Germany.
Genes Dev. 1999 May 1;13(9):1140-55. doi: 10.1101/gad.13.9.1140.
We have established a one-hybrid screen that allows the in vivo localization of proteins at a functional Saccharomyces cerevisiae centromere. Applying this screen we have identified three proteins-Ctf19, Mcm21, and the product of an unspecified open reading frame that we named Okp1-as components of the budding yeast centromere. Ctf19, Mcm21, and Okp1 most likely form a protein complex that links CBF3, a protein complex directly associated with the CDE III element of the centromere DNA, with further components of the budding yeast centromere, Cbf1, Mif2, and Cse4. We demonstrate that the CDE III element is essential and sufficient to localize the established protein network to the centromere and propose that the interaction of the CDE II element with the CDE III localized protein complex facilitates a protein-DNA conformation that evokes the active centromere.
我们建立了一种单杂交筛选方法，可在体内对酿酒酵母功能着丝粒处的蛋白质进行定位。应用此筛选方法，我们鉴定出三种蛋白质——Ctf19、Mcm21以及一个未指定的开放阅读框的产物（我们将其命名为Okp1）——作为出芽酵母着丝粒的组成部分。Ctf19、Mcm21和Okp1很可能形成一种蛋白质复合物，该复合物将CBF3（一种直接与着丝粒DNA的CDE III元件相关的蛋白质复合物）与出芽酵母着丝粒的其他组成部分Cbf1、Mif2和Cse4连接起来。我们证明，CDE III元件对于将已建立的蛋白质网络定位到着丝粒至关重要且足够，并且提出CDE II元件与CDE III定位的蛋白质复合物之间的相互作用促进了一种蛋白质 - DNA构象，从而引发活性着丝粒。
