title: miRNA proxy approach reveals hidden functions of glycosylation.
abstract: Glycosylation, the most abundant posttranslational modification, holds an unprecedented capacity for altering biological function. Our ability to harness glycosylation as a means to control biological systems is hampered by our inability to pinpoint the specific glycans and corresponding biosynthetic enzymes underlying a biological process. Herein we identify glycosylation enzymes acting as regulatory elements within a pathway using microRNA (miRNA) as a proxy. Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. Silencing these glycogenes phenocopied the effect of miR-200f, inducing mesenchymal-to-epithelial transition. In addition, all three are up-regulated in TGF-β-induced EMT, suggesting tight integration within the EMT-signaling network. Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states.
authors.affiliation: Biomedical Chemistry Institute, Department of Chemistry, New York University, New York, NY 10003.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Kurcon
authors.affiliation: Biomedical Chemistry Institute, Department of Chemistry, New York University, New York, NY 10003.
authors.forename: Zhongyin
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Biomedical Chemistry Institute, Department of Chemistry, New York University, New York, NY 10003.
authors.forename: Anika V
authors.initials: AV
authors.lastname: Paradkar
authors.affiliation: Biomedical Chemistry Institute, Department of Chemistry, New York University, New York, NY 10003.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Vaiana
authors.affiliation: Biomedical Chemistry Institute, Department of Chemistry, New York University, New York, NY 10003.
authors.forename: Sujeethraj
authors.initials: S
authors.lastname: Koppolu
authors.affiliation: Biomedical Chemistry Institute, Department of Chemistry, New York University, New York, NY 10003.
authors.forename: Praveen
authors.initials: P
authors.lastname: Agrawal
authors.affiliation: Biomedical Chemistry Institute, Department of Chemistry, New York University, New York, NY 10003 lkmahal@nyu.edu.
authors.forename: Lara K
authors.initials: LK
authors.lastname: Mahal
mesh_terms: D045744:Cell Line, Tumor; D058750:Epithelial-Mesenchymal Transition; D020868:Gene Silencing; D006031:Glycosylation; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D014166:Transferases / Q000235:genetics / Q000378:metabolism
chemical_list: D035683:MicroRNAs; D014166:Transferases
keywords: TGF-beta; epithelial to mesenchymal transition; glycan regulation; glycomics; miR-200