A Genetically Encoded Assay System to Quantify O-GlcNAc Transferase (OGT) Activity in Live Cells.

ABSTRACT: O-GlcNAc transferase (OGT) catalyzes O-GlcNAcylation of many nucleocytoplasmic proteins and plays important roles in regulating diverse cellular functions. Dysregulation of OGT is implicated in various diseases, including cancers and neurodegeneration. Despite its vital roles, little is known about how this enzyme is regulated within cells in part because no current assays directly report on its activity within cells. Here we describe a genetically encoded reporter of cellular OGT glycosyltransferase activity by exploiting the transferase-dependent proteolytic activity of OGT on host cell factor-1 (HCF-1). The reporter comprises sites at which OGT cleaves HCF-1, which are flanked by two different fluorescent proteins that are linked to either nuclear export or import sequences. OGT-catalyz

SUBMITTER: Wu WB 

PROVIDER: S-EPMC12535385 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature