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首页> 外文期刊>Molecular biology of the cell >Modeling the dynamic behaviors of the COPI vesicle formation regulators, the small GTPase Arf1 and its activating Sec7 guanine nucleotide exchange factor GBF1 on Golgi membranes
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Modeling the dynamic behaviors of the COPI vesicle formation regulators, the small GTPase Arf1 and its activating Sec7 guanine nucleotide exchange factor GBF1 on Golgi membranes

机译：对Copi囊泡形成调节剂，小GTPAse ARF1的动态行为进行建模及其激活SEC7鸟嘌呤核苷酸交换因子GBF1对Golgi膜

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摘要

The components and subprocesses underlying the formation of COPI-coated vesicles at the Golgi are well understood. The coating cascade is initiated after the small GTPase Arf1 is activated by the Sec7 domain–containing guanine nucleotide exchange factor GBF1 (Golgi brefeldin A resistant guanine nucleotide exchange factor 1). This causes a conformational shift within Arf1 that facilitates stable association of Arf1 with the membrane, a process required for subsequent recruitment of the COPI coat. Although we have atomic-level knowledge of Arf1 activation by Sec7 domain–containing GEFs, our understanding of the biophysical processes regulating Arf1 and GBF1 dynamics is limited. We used fluorescence recovery after photobleaching data and kinetic Monte Carlo simulation to assess the behavior of Arf1 and GBF1 during COPI vesicle formation in live cells. Our analyses suggest that Arf1 and GBF1 associate with Golgi membranes independently, with an excess of GBF1 relative to Arf1. Furthermore, the GBF1-mediated Arf1 activation is much faster than GBF1 cycling on/off the membrane, suggesting that GBF1 is regulated by processes other than its interactions Arf1. Interestingly, modeling the behavior of the catalytically inactive GBF1/E794K mutant stabilized on the membrane is inconsistent with the formation of a stable complex between it and an endogenous Arf1 and suggests that GBF1/E794K is stabilized on the membrane independently of complex formation.
机译：在GOLGI上形成COPI涂覆的囊泡的组分和亚过程得到了很好的理解。在通过SEC7结构域的鸟嘌呤核苷酸交换因子GBF1（Golgi Brefeldin抗性鸟嘌呤核苷酸交换因子1）激活之后，开始涂层级联。这导致ARF1内的构象变化有助于ARF1与膜的稳定关联，随后募集COPI涂层所需的方法。虽然我们对含Sec7域的全球环境基金进行ARF1激活的原子级了解，但我们对调节ARF1和GBF1动态的生物物理过程的理解有限。我们使用光漂白数据和动力学蒙特卡罗模拟后使用荧光恢复，以评估活细胞中COPI囊泡形成期间ARF1和GBF1的行为。我们的分析表明，ARF1和GBF1与Golgi膜独立相关联，相对于ARF1有超过GBF1。此外，GBF1介导的ARF1活化比循环/关闭膜的GBF1快得多，表明GBF1通过其相互作用除ARF1之外的方法进行调节。有趣的是，在膜上稳定在膜上稳定的催化活性GBF1 / E794K突变体的性能与形成其与内源性ARF1之间的稳定复合物的形成不一致，并表明GBF1 / E794K独立于复杂的形成稳定在膜上。

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