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Microarray studies on O-GlcNAcase knock out mouse embryonic fibroblast (MEF) cells show effects on metabolism and cell proliferation pathways

Wednesday, October 10, 2012 — Poster Session II

Noon – 2:00 p.m

Natcher Conference Center, Building 45




  • C.N. Keembiyehetty
  • M. Comly
  • D. C. Love
  • J. A. Hanover


The O-GlcNAc modification is analogous to phosphorylation and occurs at serine and/or threonine residues of many proteins. O- GlcNAc Transferase (OGT) and O-GlcNAcase (OGA) are enzymes that modify proteins by adding and cleaving a single O-GlcNAc moiety respectively. Perturbation of O-GlcNAc cycling is associated with diabetes, Alzheimer’s disease and cancer. Ogt is an essential gene for mouse embryonic development. The present study has developed an Oga knockout MEF cells to investigate the role of the O-GlcNAc modification. Oga knockout MEF cells were generated using the cre-loxP mating technique and MEF cells were isolated at embryonic day 12.5. Oga mRNA was ablated in Oga KO MEF cells, as compared against wild type, and consequently demonstrated higher O-GlcNAcylation of proteins. Microarray analysis was performed to identify the overall effects of Oga deletion on cell functions. ‘Go enrichment’ analysis using a set of differentially expressed, significant genes with 2 fold changes, revealed association with cell proliferation, metabolic process and growth. Elevated GSK 3 beta phosphorylation was observed in Oga deleted MEF cells under basal and insulin stimulated conditions. Oga deleted MEF cells indicate a slower proliferation rate. These findings demonstrate metabolic defects associated with altered O-GlcNAc cycling.

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