Cellular functions of the Golgi are determined by unique distribution its resident proteins. Currently, electron microscopy is required for localization a protein at sub-Golgi level. We developed quantitative method based on centers fluorescence masses nocodazole-induced ministacks under conventional optical microscopy. Our rapid, convenient, and quantitative, it yields practical resolution ∼30 nm. The was validated previous data. quantitatively studied intra-Golgi trafficking synchronized...

Proteins are transported among eukaryotic organelles along the cytoskeleton in membrane carriers. The mechanism regarding motility of carriers and positioning is a fundamental question cell biology that remains incompletely understood. Here, we find Dopey1 Mon2 assemble into complex localize to Golgi, endolysosome endoplasmic reticulum exit site. Golgi localization requires their binding phosphatidylinositol-4-phosphate phosphatidic acid, respectively, two lipids known for biogenesis...

Most proteins in the secretory pathway are glycosylated. However, role of glycans membrane trafficking is still unclear. Here, we discovered that transmembrane cargos, such as interleukin 2 receptor α subunit or Tac, transferrin receptor, and cluster differentiation 8a, unexpectedly displayed substantial Golgi localization when their O-glycosylation was compromised. By quantitatively measuring residence times, found observed O-glycan–deficient cargos due to slow export. Using a...

The endosome-to-Golgi or endocytic retrograde trafficking pathway is an important post-Golgi recycling route. Here we show that amino acids (AAs) can stimulate the and regulate cell surface localization of certain Golgi membrane proteins. By testing components AA-stimulated mTORC1 signaling pathway, demonstrate SLC38A9, v-ATPase Ragulator, but not Rag GTPases mTORC1, are essential for trafficking. Arl5, ARF-like family small GTPase, interacts with Ragulator in AA-regulated manner both Arl5...

How Golgi glycosyltransferases and glycosidases (hereafter glycosyltransferases) localize to the is still unclear. Here, we first investigated post-Golgi trafficking of glycosyltransferases. We found that can escape plasma membrane, where they are subsequently endocytosed endolysosome. Post-Golgi probably degraded by ectodomain shedding. discovered most not retrieved from sites, indicating retention rather than retrieval primary mechanism for their localization. therefore used residence time...

How the intra-Golgi secretory transport works remains a mystery. The cisternal progression and stable compartment models have been proposed are under debate. Classic model posits that both Golgi exit of cargos should occur at constant velocity dictated by progression; furthermore, COPI-mediated retrograde is essential for maintaining organization. Leveraging our recently developed imaging tools in nocodazole-induced ministacks, we found cargo decreases during their transition from cis to...

The Golgi complex consists of serially stacked membrane cisternae which can be further categorized into sub-Golgi regions, including the cis-Golgi, medial-Golgi, trans-Golgi and network. Cellular functions are determined by characteristic distribution its resident proteins. spatial resolution conventional light microscopy is too low to resolve structure or cisternae. Thus, immuno-gold electron a method choice localize protein at level. However, technique instrument beyond capability most...

Neural circuits develop through a plastic phase orchestrated by genetic programs and environmental signals. We have identified leucine-rich-repeat domain transmembrane protein PAN-1 as factor required for synaptic rewiring in

The Golgi complex consists of serially stacked membrane cisternae which can be further categorized into sub-Golgi regions, including the cis-Golgi, medial-Golgi, trans-Golgi and network. Cellular functions are determined by characteristic distribution its resident proteins. spatial resolution conventional light microscopy is too low to resolve structure or cisternae. Thus, immuno-gold electron a method choice localize protein at level. However, technique instrument beyond capability most...

ABSTRACT How Golgi glycosyltransferases and glycosidases (hereafter glycosyltransferases) localize to the is still unclear. Here, we first investigated post-Golgi trafficking of glycosyltransferases. We found that can escape plasma membrane, where they are subsequently endocytosed endolysosome. Post-Golgi probably degraded by ecto-domain shedding. discovered most not retrieved from sites, indicating retention but retrieval should be main mechanism for their localization. proposed use...

Abstract How the intra-Golgi secretory transport works remains a mystery. The cisternal progression and stable compartment models have been proposed are under debate. Classic model posits that both Golgi exit of cargos should occur at constant velocity dictated by progression; furthermore, COPI-mediated retrograde is essential for maintaining organization. Leveraging our recently developed imaging tools in nocodazole-induced ministacks, we found cargo decreases during their transition from...

How the intra-Golgi secretory transport works remains a mystery. The cisternal progression and stable compartment models have been proposed are under debate. Classic model posits that both Golgi exit of cargos should occur at constant velocity dictated by progression; furthermore, COPI-mediated retrograde is essential for maintaining organization. Leveraging our recently developed imaging tools in nocodazole-induced ministacks, we found cargo decreases during their transition from cis to...

Abstract Most proteins in the secretory pathway are glycosylated. However, role of glycans membrane trafficking is still unclear. Here, we discovered that transmembrane cargos, such as interleukin 2 receptor α subunit or Tac, transferrin and cluster differentiation 8a, unexpectedly displayed substantial Golgi localization when their O-glycosylation was compromised. By quantitatively measuring residence times, found apparent these O-glycan deficient cargos due to slow export. The...

Abstract The endosome-to-Golgi or endocytic retrograde trafficking pathway is an important post-Golgi recycling route. We made a novel discovery that the of cargos inhibited and stimulated by absence presence, respectively, amino acids (AAs), especially glutamine. By testing components AA-stimulated mTORC1 signaling pathway, we demonstrated SLC38A9, v-ATPase Ragulator, but not Rag GTPases mTORC1, are essential for trafficking. Arl5, ARF-like family small GTPase, interacts with Ragulator in...