期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2015
卷号：112
期号：23
页码：7327-7332
DOI：10.1073/pnas.1502076112
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：SignificanceCarbohydrates hold an unprecedented capacity for altering biological function, but determining which glycans and underlying enzymes are crucial for a specific biological pathway is a major impediment to our understanding of this posttranslational modification. Here we demonstrate that the mRNA target networks of microRNA (miRNA), small noncoding RNA, identify glycosylation enzymes acting as regulatory elements within a biological pathway. Leveraging the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we identify multiple promesenchymal glycosylation enzymes. Silencing miR-200f-targeted glycogenes phenocopies the effect of miR-200f, inducing mesenchymal-to-epithelial transition. These enzymes are upregulated in TGF-{beta}-induced EMT, suggesting tight integration within the signaling network. Our work indicates that miRNA networks can be used to identify crucial glycosylation enzymes driving disease states. 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-{beta}-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.
