OGT Regulates Cell Adhesion By O-GlcNAclation Of ZYX

Background and objectiveProtein post-translational modification plays an important role in many cell processes.O-GlcNAcylation modification is an important protein post-translational modifications and functions in a variety of cellular processes,such as m RNA transcription,cell cycle rergulation,neurodevelopment,stress response and metabolic homeostasis.It has been proved to be involved in neurodegenerative diseases and cancer.OGT is the O-GlcNAc transferase in O-GlcNAcylation modification.Focal adhesion is a special cell adhesion structure which connects extracellular matrix and cytoskeleton and regulates cell adhesion and migration.ZYX protein is an important component of focal adhesion and involves in the fomation of focal adhesion.This study explored the molecular mechanism by which OGT regulates cell adhesion and migration and found that OGT regulates cell adhesion and migration by O-GlcNAcylation of focal adhesion protein ZYX.MethodThe protein expression level of OGT was reduced by RNA interference technology,the interaction between OGT and ZYX was detected by co-immunoprecipitation experiments,O-GlcNAcylation of ZYX protein were detected by western blot;the stable cell line of sh RNA and sg RNA were established by RNAi and CRISPR technology respectively,the effects of OGT and O-GlcNAcylation of ZYX on cell migration were detected by cell scratch assay.ResultWe found that O-GlcNAc modification and OGT were involved in the regulation of cell adhesion and migration.ZYX,an important component in focal adhesion,was identified as a substrate of OGT.Furthermore,we found that ZYX was O-GlcNAcylated at Ser169 and Ser246 by OGT.OGT-mediated O-GlcNAcylation of ZYX was important for cell adhesion and migration.ConclusionIn this study,ZYX was found as a novel substrate of OGT.We also indentified Ser169 and Ser246 as the major O-GlcNAcylation sites of ZYX.Finally,we uncover that OGT could regulate cell adhesion and migration through O-GlcNAcylation of ZYX at Ser169 and Ser246.