A genome-wide CRISPR screen reconciles the role of N-linked glycosylation in galectin-3 transport to the cell surface.
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Authors
Menzies, Sam A
Savinykh, Natalia
Petrunkina Harrison, Anna
Publication Date
2017-10-01Journal Title
J Cell Sci
ISSN
0021-9533
Publisher
The Company of Biologists
Volume
130
Issue
19
Pages
3234-3247
Language
eng
Type
Article
This Version
AM
Physical Medium
Print-Electronic
Metadata
Show full item recordCitation
Stewart, S., Menzies, S. A., Popa, S., Savinykh, N., Petrunkina Harrison, A., Lehner, P., & Moreau, K. (2017). A genome-wide CRISPR screen reconciles the role of N-linked glycosylation in galectin-3 transport to the cell surface.. J Cell Sci, 130 (19), 3234-3247. https://doi.org/10.1242/jcs.206425
Abstract
Galectins are a family of lectin binding proteins expressed both intracellularly and extracellularly. Galectin-3 (Gal-3, also known as LGALS3) is expressed at the cell surface; however, Gal-3 lacks a signal sequence, and the mechanism of Gal-3 transport to the cell surface remains poorly understood. Here, using a genome-wide CRISPR/Cas9 forward genetic screen for regulators of Gal-3 cell surface localization, we identified genes encoding glycoproteins, enzymes involved in N-linked glycosylation, regulators of ER-Golgi trafficking and proteins involved in immunity. The results of this screening approach led us to address the controversial role of N-linked glycosylation in the transport of Gal-3 to the cell surface. We find that N-linked glycoprotein maturation is not required for Gal-3 transport from the cytosol to the extracellular space, but is important for cell surface binding. Additionally, secreted Gal-3 is predominantly free and not packaged into extracellular vesicles. These data support a secretion pathway independent of N-linked glycoproteins and extracellular vesicles.
Keywords
Hela Cells, Endoplasmic Reticulum, Golgi Apparatus, Humans, Galectin 3, Protein Transport, Glycosylation, Genome-Wide Association Study, CRISPR-Cas Systems
Sponsorship
Medical Research Council (MC_UU_12012/5)
Medical Research Council (MC_UU_12012/1)
Wellcome Trust (101835/Z/13/Z)
Wellcome Trust (100574/Z/12/Z)
Identifiers
External DOI: https://doi.org/10.1242/jcs.206425
This record's URL: https://www.repository.cam.ac.uk/handle/1810/270047
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