Sreenivasan A, Kellogg D
Sinsheimer Laboratories, Department of Biology, University of California, Santa Cruz, California 95064, USA.
Mol Cell Biol. 1999 Dec;19(12):7983-94. doi: 10.1128/MCB.19.12.7983.
In budding yeast, the Clb2 mitotic cyclin initiates a signaling network that negatively regulates polar bud growth during mitosis. This signaling network appears to require the function of a Clb2-binding protein called Nap1, the Cdc42 GTPase, and two protein kinases called Gin4 and Cla4. In this study, we demonstrate that the Elm1 kinase also plays a role in the control of bud growth during mitosis. Cells carrying a deletion of the ELM1 gene undergo a prolonged mitotic delay, fail to negatively regulate polar bud growth during mitosis, and show defects in septin organization. In addition, Elm1 is required in vivo for the proper regulation of both the Cla4 and Gin4 kinases and interacts genetically with Cla4, Gin4, and the mitotic cyclins. Previous studies have suggested that Elm1 may function to negatively regulate the Swe1 kinase. To further understand the functional relationship between Elm1 and Swe1, we have characterized the phenotype of Deltaelm1 Deltaswe1 cells. We found that Deltaelm1 Deltaswe1 cells are inviable at 37 degrees C and that a large proportion of Deltaelm1 Deltaswe1 cells grown at 30 degrees C contain multiple nuclei, suggesting severe defects in cytokinesis. In addition, we found that Elm1 is required for the normal hyperphosphorylation of Swe1 during mitosis. We propose a model in which the Elm1 kinase functions in a mitotic signaling network that controls events required for normal bud growth and cytokinesis, while the Swe1 kinase functions in a checkpoint pathway that delays nuclear division in response to defects in these events.
在芽殖酵母中,Clb2有丝分裂周期蛋白启动一个信号网络,该网络在有丝分裂期间对极性芽生长进行负调控。这个信号网络似乎需要一种名为Nap1的Clb2结合蛋白、Cdc42 GTP酶以及两种名为Gin4和Cla4的蛋白激酶发挥作用。在本研究中,我们证明Elm1激酶在有丝分裂期间对芽生长的控制中也发挥作用。携带ELM1基因缺失的细胞会经历长时间的有丝分裂延迟,在有丝分裂期间无法对极性芽生长进行负调控,并且在隔膜组织上表现出缺陷。此外,体内实验表明,Elm1对于Cla4和Gin4激酶的正常调控是必需的,并且在遗传上与Cla4、Gin4以及有丝分裂周期蛋白相互作用。先前的研究表明,Elm1可能起到对Swe1激酶进行负调控的作用。为了进一步了解Elm1和Swe1之间的功能关系,我们对Δelm1Δswe1细胞的表型进行了表征。我们发现,Δelm1Δswe1细胞在37℃时无法存活,并且在30℃下生长的大部分Δelm1Δswe1细胞含有多个细胞核,这表明在胞质分裂方面存在严重缺陷。此外,我们发现有丝分裂期间Swe1的正常过度磷酸化需要Elm1。我们提出了一个模型,其中Elm1激酶在一个有丝分裂信号网络中发挥作用,该网络控制正常芽生长和胞质分裂所需的事件,而Swe1激酶在一个检查点途径中发挥作用,该途径会响应这些事件中的缺陷而延迟核分裂。