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Dissection of mitotic functions of the yeast cyclin Clb2

机译：剖析酵母细胞周期蛋白Clb2的有丝分裂功能

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摘要

Progression through mitosis requires the activity of cyclin-dependent kinases (CDKs) associated with regulatory cyclin subunits. In the yeast Saccharomyces cerevisiae, Clb2 has the most important role among the four mitotic cyclins, Clb1-4, manifested by data showing that simultaneous deletion of the CLB1, CLB3 and CLB4 genes has only minor effects on mitosis. Thus, Clb2 alone is sufficient for all essential CDK functions in mitosis, such as the assembly of bipolar spindles and spindle elongation. Here, we show that a modification of Clb2, by the C-terminal addition of a Myc12 epitope, causes the loss of one specific mitotic function of Clb2. Strains carrying CLB2-MYC12 are nonviable in the absence of the CLB3 and CLB4 genes, because the modified Clb2 version fails to promote assembly of the mitotic spindle. In contrast, Clb2-Myc12 has no apparent defects in late mitotic functions and, furthermore, induces the switch from polarized to isotropic growth with similar efficiency as the endogenous Clb2. Thus, the presence of the Myc12 epitope selectively inactivates Clb2's capacity to promote spindle formation. Clb2-Myc12 represents therefore the first version of Clb2 impaired in one specific mitotic function. We conclude that the major mitotic functions of this cyclin can be unequivocally dissected.
机译：通过有丝分裂的进展需要与调节性细胞周期蛋白亚基相关的细胞周期蛋白依赖性激酶（CDK）的活性。在酵母酿酒酵母中，Clb2在四个有丝分裂细胞周期蛋白Clb1-4中具有最重要的作用，其数据表明同时删除CLB1，CLB3和CLB4基因对有丝分裂的影响很小。因此，单独的Clb2足以满足有丝分裂中所有重要的CDK功能，例如双极纺锤体的组装和纺锤体伸长。在这里，我们表明，Cb2的修饰，通过Myc12表位的C末端添加，会导致Clb2的一种特定的有丝分裂功能丧失。在缺少CLB3和CLB4基因的情况下，携带CLB2-MYC12的菌株是不可行的，因为修饰的Clb2版本无法促进有丝分裂纺锤体的组装。相反，Clb2-Myc12在后期的有丝分裂功能上没有明显的缺陷，而且还以与内源性Clb2相似的效率诱导了从极化生长向各向同性生长的转变。因此，Myc12表位的存在选择性地失活了Clb2促进纺锤体形成的能力。因此，Clb2-Myc12代表一个特定的有丝分裂功能受损的Clb2的第一个版本。我们得出结论，该细胞周期蛋白的主要有丝分裂功能可以明确地解剖。

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