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ESCargo: a regulatable fluorescent secretory cargo for diverse model organisms
Jason C Casler1, Allison L Zajac1, Fernando M Valbuena1
1Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637.
Molecular Biology of the Cell
|October 28, 2020
Summary
We developed ESCargo, a novel fluorescent protein for studying membrane traffic. This tool enables synchronized cargo waves from the endoplasmic reticulum (ER) in multiple model organisms.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Studying membrane traffic requires tools to track cargo proteins through the secretory pathway.
- Existing methods for generating synchronized cargo waves often have limitations in usability and organism specificity.
Purpose of the Study:
- To develop a versatile and easy-to-use regulatable secretory cargo for studying membrane traffic.
- To demonstrate the utility of this new tool across diverse model organisms.
Main Methods:
- Optimization of an artificial fluorescent secretory protein (ESCargo) for budding yeast.
- Utilizing the Erv29 cargo receptor (homologous to mammalian Surf4) for ER export.
- Inducing synchronized cargo waves by disaggregating ER-localized ESCargo with a ligand.
Main Results:
- ESCargo forms aggregates in the ER lumen, allowing for controlled release.
- Demonstrated successful application of ESCargo in yeast, mammalian, Drosophila, and Tetrahymena cells.
- Kinetic analysis confirmed that Erv29/Surf4 recognition is crucial for rapid ER export.
Conclusions:
- ESCargo provides a simple and effective method for generating synchronized secretory cargo waves.
- The technology is adaptable to various model organisms, facilitating broader research in membrane traffic.
- Recognition by Erv29/Surf4 is a key determinant of efficient ER export for this cargo system.
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