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首页> 外文学位 >Function and specificity in signaling pathway of two map kinases ERK1 and ERK2 in Dictyostelium.
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Function and specificity in signaling pathway of two map kinases ERK1 and ERK2 in Dictyostelium.

机译：盘基网柄菌中两个图激酶ERK1和ERK2在信号传导途径中的功能和特异性。

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摘要

Scope and Method of Study: MAPKs are associated with several human diseases. However, studying MAPK signaling pathways in higher eukaryotes is challenging due to presence of several MAPKs. Dictyostelium, whose genome encodes only two MAPKs ERK1 and ERK2, therefore offers many advantages in this respect. Our purpose in this research is to define the function and the specificity in signaling pathway of ERK1 and ERK2 through characterizing G protein-mediated pathways and searching for ERK-associated proteins. We used immunoprecipitation, pull-down assays and genetic analyses to examine the interactions of ERKs with wild-type and mutated Ga subunits. We also created ERK mutants and performed the complementation, suppression and epistasis tests to determine role of ERKs in Ga subunit-mediated pathways. To search for ERK-associated proteins, His6-tagged ERK1 and ERK2 complexes were isolated and then subjected to mass spectrometry analysis. Cellular distribution of the transcription factors (STATs), expression of the cell type-specific genes (ecmA, ecmB) and the knockout of the phosphodiesterase RegA were used to determine the involvement of these ERK-associated proteins in ERK pathways.;Findings and Conclusions: Our results showed that the Ga4 subunit interacts with ERK2 and the Ga5 subunit associates with ERK1 through the D-motifs. The Ga4-ERK2 interaction contributes to spore development while the Ga5-ERK1 interaction regulates cell viability and tip morphogenesis. We also indicated that ERK1 and ERK2 have different roles in Ga subunit-mediated pathways. Genetic analyses of the ERK-associated proteins revealed that both ERK1 and ERK2 negatively regulate RegA; the Ga5 subunit-ERK1 pathway controls nuclear localization of STATc ultimately resulting in repression of ecmA while the Ga2/Ga4 subunits-ERK2 pathways regulate function of STATa and expression of ecmB through down regulating RegA.
机译：研究范围和方法：MAPK与几种人类疾病有关。但是，由于存在几种MAPK，因此在高等真核生物中研究MAPK信号通路具有挑战性。因此，其基因组仅编码两个MAPK ERK1和ERK2的双壁铁锈菌在这方面具有许多优势。我们在这项研究中的目的是通过表征G蛋白介导的途径并寻找ERK相关蛋白来定义ERK1和ERK2信号通路中的功能和特异性。我们使用免疫沉淀，下拉测定法和遗传分析来检查ERKs与野生型和突变的Ga亚基的相互作用。我们还创建了ERK突变体，并进行了互补，抑制和上位性测试，以确定ERK在Ga亚基介导的途径中的作用。为了搜索与ERK相关的蛋白质，分离了带有His6标签的ERK1和ERK2复合物，然后进行质谱分析。转录因子（STATs）的细胞分布，细胞类型特异性基因（ecmA，ecmB）的表达以及磷酸二酯酶RegA的敲除被用来确定这些ERK相关蛋白在ERK途径中的参与。;发现和结论：我们的结果显示，Ga4亚基与ERK2相互作用，而Ga5亚基通过D-基元与ERK1缔合。 Ga4-ERK2相互作用有助于孢子发育，而Ga5-ERK1相互作用则调节细胞活力和尖端形态发生。我们还表明，ERK1和ERK2在Ga亚基介导的途径中具有不同的作用。对ERK相关蛋白的遗传分析表明，ERK1和ERK2均对RegA产生负调控。 Ga5亚基-ERK1通路控制STATc的核定位，最终导致ecmA的抑制，而Ga2 / Ga4亚基-ERK2通路通过下调RegA调节STATa的功能和ecmB的表达。

著录项

  • 作者

    Nguyen, Hoai-Nghia.;

  • 作者单位

    Oklahoma State University.;

  • 授予单位 Oklahoma State University.;
  • 学科 Biology Cell.;Health Sciences Human Development.
  • 学位 Ph.D.
  • 年度 2011
  • 页码 156 p.
  • 总页数 156
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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