A Set of Fluorescent Protein-Based Markers for Major Vesicle Coat Proteins in Yeast
Xue-Fei Cui, Zheng-Tan Zhang, Jing Zhu, Li Cui, Zhiping Xie
TL;DR
This paper introduces fluorescent markers for coat proteins in yeast, revealing their locations and providing evidence for transport vesicles.
Contribution
The paper introduces a novel set of fluorescent markers for major vesicle coat proteins in yeast.
Findings
COPII puncta were primarily associated with the endoplasmic reticulum and occasionally with early Golgi.
AP-1 and AP-3 puncta were often found not at their donor compartments, suggesting they are coated transport vesicles.
Retromer was localized to vacuoles, late endosomes, and perivacuolar puncta.
Abstract
In eukaryotic cells, vesicle-mediated transport interconnects the endomembrane system. These vesicles are formed by coat proteins via deformation of donor membranes. Here, we constructed a set of fluorescent protein-based markers for major coat protein complexes in the yeast model system, and examined their subcellular localization patterns. Our markers covered COPII, COPI, AP-1, AP-2, AP-3, and retromer complexes. Our live cell imaging demonstrates that COPII puncta were primarily associated with the endoplasmic reticulum (ER), and occasionally with early Golgi. COPI was present on both early Golgi and late Golgi/early endosomes. AP-1 puncta were present on late Golgi/early endosomes. AP-2 was present on plasma membrane (PM)-associated puncta, and around the bud neck. AP-3 puncta were present on late Golgi/early endosomes and on the surface of vacuoles. Retromer was present on the…
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Taxonomy
TopicsCellular transport and secretion · Fungal and yeast genetics research · Endoplasmic Reticulum Stress and Disease