Wysocka Monika, Rytka Joanna, Kurlandzka Anna
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02-106 Warsaw, Poland.
Exp Cell Res. 2004 Apr 1;294(2):592-602. doi: 10.1016/j.yexcr.2003.12.008.
We present the characteristics of the Csm1 (Spo86) protein of Saccharomyces cerevisiae that are important for meiotic division. The level of Csm1p does not change throughout the cell cycle, but this protein is absent in mature spores. Deletion of CSM1 causes incorrect spore formation and meiotic chromosome missegregation together with increased sensitivity of vegetative cells to benomyl and manganese. In a two-hybrid analysis with Csm1p as bait, we detected interactions with three members of the Mcm2-7 family of proteins involved in the initiation of DNA replication, and with Clf1p also implicated in replication. The Csm1p-Mcm3, Mcm5 and Mcm7p interactions were confirmed by co-immunoprecipitation. Three other interacting proteins, Mgs1p, Ulp2, and Plp2, participate in chromosome assembling and segregation, whereas the function of two others has not been established. Genetic experiments showed that the two-hybrid isolates MGS1, CLF1, MCM3, 5, 7 (CDC47), and YDL089w, when overexpressed, partially suppress the csm1Delta/csm1Delta sporulation defect. We propose that, besides its other functions, Csm1p may be involved in premeiotic DNA replication.
我们展示了酿酒酵母中对减数分裂至关重要的Csm1(Spo86)蛋白的特征。Csm1p的水平在整个细胞周期中都不会改变,但这种蛋白在成熟孢子中不存在。CSM1的缺失会导致孢子形成错误和减数分裂染色体错配,同时营养细胞对苯菌灵和锰的敏感性增加。在以Csm1p为诱饵的双杂交分析中,我们检测到它与参与DNA复制起始的Mcm2 - 7蛋白家族的三个成员以及也与复制有关的Clf1p相互作用。通过共免疫沉淀证实了Csm1p与Mcm3、Mcm5和Mcm7p的相互作用。其他三个相互作用蛋白,Mgs1p、Ulp2和Plp2,参与染色体组装和分离,而另外两个蛋白的功能尚未确定。遗传实验表明,双杂交分离出的MGS1、CLF1、MCM3、5、7(CDC47)和YDL089w在过表达时,部分抑制了csm1Delta/csm1Delta孢子形成缺陷。我们提出,除了其其他功能外,Csm1p可能参与减数分裂前的DNA复制。