Inhibition of cargo export at ER exit sites and the trans-Golgi network by the secretion inhibitor FLI-06

Yonemura, Yoji; Li, Xin; Müller, Katja; Krämer, Andreas; Atigbire, Paul; Mentrup, Torben; Feuerhake, Talitha; Kroll, Torsten; Shomron, Olga; Nohl, Richard; Arndt, Hans‐Dieter; Hoischen, Christian; Hemmerich, Peter; Hirschberg, Koret; Kaether, Christoph

doi:10.1242/jcs.186163

Cited by 18 publications

(31 citation statements)

References 62 publications

(80 reference statements)

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“…GCA insensitivity indicates that the NS1 and CAV-1 interaction initiates in the ER compartment. In agreement, treatment of mosquito cells with Fli-06 showed no effect on NS1 secretion, indicating that the interaction between NS1 and CCC takes place very early after NS1 synthesis, before the viral protein is transported to the ERES (22,23).…”

Section: Discussionsupporting

confidence: 56%

The Dengue Virus Nonstructural Protein 1 (NS1) Is Secreted from Mosquito Cells in Association with the Intracellular Cholesterol Transporter Chaperone Caveolin Complex

Rosales

Ludert

2019

J Virol

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Dengue virus (DENV) is a mosquito-borne virus of the family Flaviviridae. The RNA viral genome encodes three structural and seven nonstructural proteins. Nonstructural protein 1 (NS1) is a multifunctional protein actively secreted in vertebrate and mosquito cells during infection. In mosquito cells, NS1 is secreted in a caveolin-1-dependent manner by an unconventional route. The caveolin chaperone complex (CCC) is a cytoplasmic complex formed by caveolin-1 and the chaperones FKBP52, Cy40, and CyA and is responsible for the cholesterol traffic inside the cell. In this work, we demonstrate that in mosquito cells, but not in vertebrate cells, NS1 associates with and relies on the CCC for secretion. Treatment of mosquito cells with classic secretion inhibitors, such as brefeldin A, Golgicide A, and Fli-06, showed no effect on NS1 secretion but significant reductions in recombinant luciferase secretion and virion release. Silencing the expression of CAV-1 or FKBP52 with short interfering RNAs or the inhibition of CyA by cyclosporine resulted in significant decrease in NS1 secretion, again without affecting virion release. Colocalization, coimmunoprecipitation, and proximity ligation assays indicated that NS1 colocalizes and interacts with all proteins of the CCC. In addition, CAV-1 and FKBP52 expression was found augmented in DENV-infected cells. Results obtained with Zika virus-infected cells suggest that in mosquito cells, ZIKV NS1 follows the same secretory pathway as that observed for DENV NS1. These results uncover important differences in the dengue virus-cell interactions between the vertebrate host and the mosquito vector as well as novel functions for the chaperone caveolin complex. IMPORTANCE The dengue virus protein NS1 is secreted efficiently from both infected vertebrate and mosquito cells. Previously, our group reported that NS1 secretion in mosquito cells follows an unconventional secretion pathway dependent on caveolin-1. In this work, we demonstrate that in mosquito cells, but not in vertebrate cells, NS1 secretion takes place in association with the chaperone caveolin complex, a complex formed by caveolin-1 and the chaperones FKBP52, CyA, and Cy40, which are in charge of cholesterol transport inside the cell. Results obtained with ZIKV-infected mosquito cells suggest that ZIKV NS1 is released following an unconventional secretory route in association with the chaperone caveolin complex. These results uncover important differences in the virus-cell interactions between the vertebrate host and the mosquito vector, as well as novel functions for the chaperone caveolin complex. Moreover, manipulation of the NS1 secretory route may prove a valuable strategy to combat these two mosquito-borne diseases. Volume 93 Issue 4 e01985-18 jvi.asm.org 2 on March 18, 2020 by guest http://jvi.asm.org/ Downloaded from plasma membrane, mediated by the CCC, to reach the extracellular space. In addition, data are presented suggesting that a similar pathway is used for the secretion of Zika virus NS1 protein in infected m...

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Section: Discussionsupporting

confidence: 56%

The Dengue Virus Nonstructural Protein 1 (NS1) Is Secreted from Mosquito Cells in Association with the Intracellular Cholesterol Transporter Chaperone Caveolin Complex

Rosales

Ludert

2019

J Virol

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“…Upon shifting to the permissive temperature (32 °C), VSVG-EYFP folds properly, exits the ER and travels to the plasma membrane [36, 37]. Using its N-glycosylation sites the localization can be assessed with the help of endoglycosidaseH (endoH), because ER-localized VSVG-EYFP is endoH-sensitive whereas Golgi and beyond-localized VSVG-EYFP is endoH resistant [31]. In HeLa cells transfected with ATL3 or ATL3 Y192C together with VSVG-EYFP only the Y192C mutation, but not the wild type, slowed down ER–Golgi transport (Fig.…”

Section: Resultsmentioning

confidence: 99%

A disease causing ATLASTIN 3 mutation affects multiple endoplasmic reticulum-related pathways

Behrendt

Kurth

Kaether

2019

Cell. Mol. Life Sci.

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Atlastins (ATLs) are membrane-bound GTPases involved in shaping of the endoplasmic reticulum (ER). Mutations in ATL1 and ATL3 cause spastic paraplegia and hereditary sensory neuropathy. We here show that the sensory neuropathy causing ATL3 Y192C mutation reduces the complexity of the tubular ER-network. ATL3 Y192C delays ER-export by reducing the number of ER exit sites, reduces autophagy, fragments the Golgi and causes malformation of the nucleus. In cultured primary neurons, ATL3 Y192C does not localize to the growing axon, resulting in axon growth deficits. Patient-derived fibroblasts possess a tubular ER with reduced complexity and have a reduced number of autophagosomes. The data suggest that the disease-causing ATL3 Y192C mutation affects multiple ER-related pathways, possibly as a consequence of the distorted ER morphology.Electronic supplementary materialThe online version of this article (10.1007/s00018-019-03010-x) contains supplementary material, which is available to authorized users.

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“…There is some scope here for further development and some early work in this area has already identified potential inhibitors of COPII-dependent budding (Yonemura et al 2016). The essential nature of the COPII pathway means that specific inhibitors are unlikely to be of therapeutic benefit; however, there might be further scope in targeting cargo-specific interactions, the interplay between protein folding and export, or the efficiency of the process itself.…”

Section: Future Perspectivesmentioning

confidence: 99%

COPII-dependent ER export in animal cells: adaptation and control for diverse cargo

McCaughey

Stephens

2018

Histochem Cell Biol

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The export of newly synthesized proteins from the endoplasmic reticulum is fundamental to the ongoing maintenance of cell and tissue structure and function. After co-translational translocation into the ER, proteins destined for downstream intracellular compartments or secretion from the cell are sorted and packaged into transport vesicles by the COPII coat protein complex. The fundamental discovery and characterization of the pathway has now been augmented by a greater understanding of the role of COPII in diverse aspects of cell function. We now have a deep understanding of how COPII contributes to the trafficking of diverse cargoes including extracellular matrix molecules, developmental signalling proteins, and key metabolic factors such as lipoproteins. Structural and functional studies have shown that the COPII coat is both highly flexible and subject to multiple modes of regulation. This has led to new discoveries defining roles of COPII in development, autophagy, and tissue organization. Many of these newly emerging features of the canonical COPII pathway are placed in a context of procollagen secretion because of the fundamental interest in how a coat complex that typically generates 80-nm transport vesicles can package a cargo reported to be over 300 nm. Here we review the current understanding of COPII and assess the current consensus on its role in packaging diverse cargo proteins.

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Inhibition of cargo export at ER exit sites and the trans-Golgi network by the secretion inhibitor FLI-06

Cited by 18 publications

References 62 publications

The Dengue Virus Nonstructural Protein 1 (NS1) Is Secreted from Mosquito Cells in Association with the Intracellular Cholesterol Transporter Chaperone Caveolin Complex

The Dengue Virus Nonstructural Protein 1 (NS1) Is Secreted from Mosquito Cells in Association with the Intracellular Cholesterol Transporter Chaperone Caveolin Complex

A disease causing ATLASTIN 3 mutation affects multiple endoplasmic reticulum-related pathways

COPII-dependent ER export in animal cells: adaptation and control for diverse cargo

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