Genetic gradual reduction of OGT activity unveils the essential role of O-GlcNAc in the mouse embryo

The reversible glycosylation of nuclear and cytoplasmic proteins (O-GlcNAcylation) is catalyzed by a single enzyme, namely O-GlcNAc transferase (OGT). The mammalianOgtgene is X-linked and it is essential for embryonic development and for the viability of proliferating cells. We perturbed OGT’s functionin vivoby creating a murine allelic series of four single amino acid substitutions reducing OGT’s catalytic activity to a range of degrees. The severity of the embryonic lethality was proportional to the degree of impairment of OGT’s catalysis, demonstrating that the O-GlcNAc modification itself is required for early development. We identified milder hypomorphicOgtalleles that perturb O-GlcNAc homeostasis while being compatible with embryogenesis. The analysis of the transcriptomes of the mutant embryos at different stages suggested a sexually-dimorphic developmental delay caused by the decrease in O-GlcNAc. Furthermore, a mild reduction of OGT’s enzymatic activity was sufficient to loosen the silencing of endogenous retrovirusesin vivo.