Live Cell Super-Resolution Imaging Research Team, RIKEN Center for Advanced Photonics, Wako, Japan.
Laboratory of Molecular Cell Biology, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan.
Elife. 2024 Mar 19;13:e92900. doi: 10.7554/eLife.92900.
Cargo traffic through the Golgi apparatus is mediated by cisternal maturation, but it remains largely unclear how the -cisternae, the earliest Golgi sub-compartment, is generated and how the Golgi matures into the -Golgi network (TGN). Here, we use high-speed and high-resolution confocal microscopy to analyze the spatiotemporal dynamics of a diverse set of proteins that reside in and around the Golgi in budding yeast. We find many mobile punctate structures that harbor yeast counterparts of mammalian endoplasmic reticulum (ER)-Golgi intermediate compartment (ERGIC) proteins, which we term 'yeast ERGIC'. It occasionally exhibits approach and contact behavior toward the ER exit sites and gradually matures into the -Golgi. Upon treatment with the Golgi-disrupting agent brefeldin A, the ERGIC proteins form larger aggregates corresponding to the Golgi entry core compartment in plants, while - and medial-Golgi proteins are absorbed into the ER. We further analyze the dynamics of several late Golgi proteins to better understand the Golgi-TGN transition. Together with our previous studies, we demonstrate a detailed spatiotemporal profile of the entire cisternal maturation process from the ERGIC to the Golgi and further to the TGN.
通过高尔基器的货物运输是通过小泡成熟介导的，但 - 潴泡（最早的高尔基亚区室）是如何产生的，以及高尔基器如何成熟为 - 高尔基网络（TGN），这在很大程度上仍不清楚。在这里，我们使用高速和高分辨率共聚焦显微镜分析了在出芽酵母中位于高尔基器内部和周围的多种蛋白质的时空动力学。我们发现了许多可移动的点状结构，其中含有酵母内质网-高尔基中间区室（ERGIC）蛋白的哺乳动物对应物，我们将其称为“酵母 ERGIC”。它偶尔会表现出接近和接触内质网出口部位的行为，并逐渐成熟为 - 高尔基器。在用破坏高尔基体的试剂布雷菲德菌素 A 处理后，ERGIC 蛋白形成与植物中的高尔基体进入核心区室相对应的更大聚集体，而 - 和中间高尔基蛋白则被吸收到内质网中。我们进一步分析了几种晚期高尔基蛋白的动力学，以更好地理解高尔基器-TGN 转变。结合我们以前的研究，我们展示了从 ERGIC 到高尔基器再到 TGN 的整个小泡成熟过程的详细时空特征。
