Language
English
Publication Date
9-15-2026
Journal
Journal of Biological Chemistry
DOI
10.1016/j.jbc.2026.113556
PMID
42744251
PMCID
PMC13683433
PubMedCentral® Posted Date
9-15-2026
PubMedCentral® Full Text Version
Post-print
Abstract
Protein O-glucosyltransferase 1 (POGLUT1/Rumi) transfers an O-linked glucose (O-Glc) monosaccharide from UDP-Glc to a serine residue in epidermal growth factor (EGF)-like repeats with a specific consensus sequence. In mammals, GXYLT1 and GXYLT2 transfer xylose to this O-Glc monosaccharide. Previous studies showed that Poglut1 deletion in mice leads to embryonic lethality with defects in crumbs family member 2 (CRB2) trafficking and NOTCH1 signaling. Both CRB2 and Notch receptors have multiple EGF repeats with the O-Glc modification site. However, the roles of xylosyl elongation of O-Glc in protein trafficking and mammalian embryonic development are unknown. Here, we demonstrated that xylosyl elongation of O-Glc occurs in the endoplasmic reticulum and comprehensively analyzed O-Glc glycosylation on CRB2, NOTCH1, and NOTCH2 expressed in HEK293T cells by mass spectrometry. Most EGF repeats containing the consensus sequence were modified with O-Glc glycans, while xylosyl elongation varied among different EGF repeats, suggesting EGF repeat-specific regulation. GXYLT1 KO cells showed reduced or lost xylosyl elongation, but GXYLT2 KO cells did not. Cell-based assays demonstrated decreased secretion of substrate proteins in GXYLT1 KO cells, which was rescued by WT GXYLT1. Analysis of Gxylt1 and Gxylt2 KO mice revealed that Gxylt1 deletion, but not Gxylt2 deletion, caused embryonic lethality. Altogether, our data suggest that GXYLT1 plays a major role in xylosyl elongation of O-Glc glycans and thereby contributes to the endoplasmic reticulum quality control and trafficking of CRB2, NOTCH1, and NOTCH2 in HEK293T cells while GXYLT1 and GXYLT2 have distinct biological functions during mouse embryonic development.
Keywords
EGF repeats, endoplasmic reticulum, ER quality control, glycosylation, glycosyltransferase, mass spectrometry, crumbs, Notch, O-glucosylation, posttranslational modification, xylose
Published Open-Access
yes
Recommended Citation
Saiki, Wataru; Okamura, Mio; Fujita, Yuki; et al., "ER Quality Control of CRUMBS2 and Notch Proteins by GXYLT1-Dependent Xylosyl Elongation of O-Glucose Glycans" (2026). Faculty, Staff and Students Publications. 8579.
https://digitalcommons.library.tmc.edu/baylor_docs/8579