Cytosolic UDP-Gal biosynthetic machinery is required for dimerization of SLC35A2 in the Golgi membrane and its interaction with B4GalT1.

ABSTRACT: Glycosylation is a vital post-translational modification involving the addition of sugars to proteins and lipids, facilitated by glycosyltransferases and dependent on nucleotide sugar donors like UDP-galactose (UDP-Gal). This study examines how disruptions in UDP-Gal synthesis affect protein-protein interactions critical for glycosylation. Using CRISPR/Cas9, we generated HEK293T cell lines lacking key enzymes of the Leloir pathway: UDP-galactose 4'-epimerase (GALE), galactose-1-phosphate uridylyltransferase (GALT), or both. The knockout of GALE led to a significant reduction in intracellular UDP-Gal levels and altered N-glycan profiles, indicating impaired galactosylation. Through the NanoBiT assay, we observed that knocking out GALE alone or both GALE and GALT diminished the ability of th

SUBMITTER: Wiertelak W 

PROVIDER: S-EPMC11994309 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature