Yeast Physiology Group, Department of Genetics and Microbiology, Facultad de Biología. Universidad de Murcia, 30071 Murcia, Spain.
J Biol Chem. 2012 Jul 27;287(31):26038-51. doi: 10.1074/jbc.M112.345611. Epub 2012 Jun 8.
Mitogen-activated protein kinase (MAPK) signaling pathways play a fundamental role in the response of eukaryotic cells to environmental changes. Also, much evidence shows that the stimulus-dependent nuclear targeting of this class of regulatory kinases is crucial for adequate regulation of distinct cellular events. In the fission yeast Schizosaccharomyces pombe, the cell integrity MAPK pathway, whose central element is the MAPK Pmk1, regulates multiple processes such as cell wall integrity, vacuole fusion, cytokinesis, and ionic homeostasis. In non-stressed cells Pmk1 is constitutively localized in both cytoplasm and nucleus, and its localization pattern appears unaffected by its activation status or in response to stress, thus questioning the biological significance of the presence of this MAPK into the nucleus. We have addressed this issue by characterizing mutants expressing Pmk1 versions excluded from the cell nucleus and anchored to the plasma membrane in different genetic backgrounds. Although nuclear Pmk1 partially regulates cell wall integrity at a transcriptional level, membrane-tethered Pmk1 performs many of the biological functions assigned to wild type MAPK like regulation of chloride homeostasis, vacuole fusion, and cellular separation. However, we found that down-regulation of nuclear Pmk1 by MAPK phosphatases induced by the stress activated protein kinase pathway is important for the fine modulation of extranuclear Pmk1 activity. These results highlight the importance of the control of MAPK activity at subcellular level.
丝裂原活化蛋白激酶(MAPK)信号通路在真核细胞对环境变化的反应中起着至关重要的作用。此外,大量证据表明,这一类调节激酶的刺激依赖性核定位对于适当调节不同的细胞事件至关重要。在裂殖酵母 Schizosaccharomyces pombe 中,细胞完整性 MAPK 途径的核心元件是 MAPK Pmk1,它调节多种过程,如细胞壁完整性、液泡融合、胞质分裂和离子动态平衡。在非应激细胞中,Pmk1 持续定位于细胞质和细胞核中,其定位模式似乎不受其激活状态或应激反应的影响,这就质疑了这种 MAPK 存在于细胞核中的生物学意义。我们通过对表达排除在细胞核之外并锚定在不同遗传背景下质膜上的 Pmk1 变体的突变体进行表征来解决这个问题。尽管核 Pmk1 在转录水平上部分调节细胞壁完整性,但膜结合的 Pmk1 执行许多被分配给野生型 MAPK 的生物学功能,如氯离子动态平衡、液泡融合和细胞分离的调节。然而,我们发现,由应激激活的蛋白激酶途径中的 MAPK 磷酸酶诱导的核 Pmk1 的下调对于核外 Pmk1 活性的精细调节很重要。这些结果强调了在亚细胞水平上控制 MAPK 活性的重要性。