MicroRNA-424 Predicts a Role for β-1,4 Branched Glycosylation in Cell Cycle Progression

J Biol Chem. 2016 Jan 15;291(3):1529-37. doi: 10.1074/jbc.M115.672220. Epub 2015 Nov 20.

Abstract

MicroRNA regulation of protein expression plays an important role in mediating many cellular processes, from cell proliferation to cell death. The human microRNA miR-424 is up-regulated in response to anti-proliferative cytokines, such as transforming growth factor β (TGFβ), and directly represses cell cycle progression. Our laboratory recently established that microRNA can be used as a proxy to identify biological roles of glycosylation enzymes (glycogenes). Herein we identify MGAT4A, OGT, and GALNT13 as targets of miR-424. We demonstrate that MGAT4A, an N-acetylglucosaminyltransferase that installs the β-1,4 branch of N-glycans, is directly regulated by miR-424 in multiple mammary epithelial cell lines and observe the loss of MGAT4A in response to TGFβ, an inducer of miR-424. Knockdown of MGAT4A induces cell cycle arrest through decreasing CCND1 levels. MGAT4A does not affect levels of β-1,6 branched N-glycans, arguing that this effect is specific to β-1,4 branching and not due to gross changes in overall N-linked glycosylation. This work provides insight into the regulation of cell cycle progression by specific N-glycan branching patterns.

Keywords: CDC25A; Galectin-3; MGAT5; N-glycan biosynthesis; N-linked glycosylation; cell cycle; complex N-glycans; cyclin D1; glycosylation; microRNA (miRNA).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle
  • Cell Line
  • Cell Proliferation
  • Cyclin D1 / antagonists & inhibitors
  • Cyclin D1 / genetics
  • Cyclin D1 / metabolism
  • Enzyme Repression
  • Genes, Reporter
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Luciferases / genetics
  • Luciferases / metabolism
  • Mammary Glands, Human / cytology
  • Mammary Glands, Human / enzymology
  • Mammary Glands, Human / metabolism*
  • MicroRNAs / agonists
  • MicroRNAs / metabolism*
  • N-Acetylgalactosaminyltransferases / antagonists & inhibitors*
  • N-Acetylgalactosaminyltransferases / genetics
  • N-Acetylgalactosaminyltransferases / metabolism
  • N-Acetylglucosaminyltransferases / antagonists & inhibitors*
  • N-Acetylglucosaminyltransferases / genetics
  • N-Acetylglucosaminyltransferases / metabolism
  • RNA Interference*
  • RNA, Small Interfering
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Transforming Growth Factor beta / metabolism

Substances

  • CCND1 protein, human
  • MIRN424 microrna, human
  • MicroRNAs
  • RNA, Small Interfering
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • Cyclin D1
  • Luciferases
  • MGAT4A protein, human
  • N-Acetylgalactosaminyltransferases
  • N-Acetylglucosaminyltransferases
  • UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase
  • GALNT13 protein, human