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Genome‐scale identification of UDP‐GlcNAc‐dependent pathways
Author(s) -
Lau Ken S.,
Khan Sofia,
Dennis James W.
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200800208
Subject(s) - microbiology and biotechnology , endocytosis , small interfering rna , glycan , biology , downregulation and upregulation , signal transduction , transfection , golgi apparatus , hek 293 cells , chemistry , cell , gene , biochemistry , glycoprotein
Metabolite flux to UDP‐GlcNAc and Golgi N ‐glycan biosynthesis regulates surface residency of glycoprotein receptors and transporters, and thus sensitivities of cells to extracellular cues. Salvage of GlcNAc increases UDP‐GlcNAc and branching of N ‐glycans progressively, but displays an optimum for cell proliferation and bulk endocytosis in mouse NMuMG and human HEK293T epithelial cells. In this report, we measured global changes in gene expression in low and high GlcNAc‐supplemented cells. Genes upregulated by high GlcNAc included the EGF and TGF‐β signaling pathways and cell cycle checkpoint, while downregulated genes indicated lower metabolic activity. Genes increased or decreased by high GlcNAc were assessed by transfecting cells with small interfering RNA (siRNA) and measuring effects on three phenotypes: proliferation and bulk endocytosis, and β1,6GlcNAc‐branching of N ‐glycans. siRNA targeting LGALS3, WBSCR17, PHF3, SDC2 and CTNNAL1 partially reversed the GlcNAc‐induced phenotypes, suggesting a role for galectin‐3/ N ‐glycans, proteoglycans, O ‐glycans, and junctional cell adhesion.