V(D)J recombination secures the production of functional immunoglobulin(Ig)genes and antibody diversity during the early stages of B-cell development through long-distance interactions mediated by cis-regulatory eleme...V(D)J recombination secures the production of functional immunoglobulin(Ig)genes and antibody diversity during the early stages of B-cell development through long-distance interactions mediated by cis-regulatory elements and trans-acting factors.O-GlcNAcylation is a dynamic and reversible posttranslational modification of nuclear and cytoplasmic proteins that regulates various protein functions,including DNA-binding affinity and protein-protein interactions.However,the effects of O-GlcNAcylation on proteins involved in V(D)J recombination remain largely unknown.To elucidate this relationship,we downregulated O-GlcNAcylation in a mouse model by administering an O-GlcNAc inhibitor or restricting the consumption of a regular diet.Interestingly,the inhibition of O-GlcNAcylation in mice severely impaired Ig heavy-chain(IgH)gene rearrangement.We identified several factors crucial for V(D)J recombination,including YY1,CTCF,SMC1,and SMC3,as direct targets of O-GlcNAc modification.Importantly,O-GlcNAcylation regulates the physical interaction between SMC1 and SMC3 and the DNA-binding patterns of YY1 at the IgH gene locus.Moreover,O-GlcNAc inhibition downregulated DDX5 protein expression,affecting the functional association of CTCF with its DNA-binding sites at the IgH locus.Our results showed that locus contraction and long-range interactions throughout the IgH locus are disrupted in a manner dependent on the cellular O-GlcNAc level.In this study,we established that V(D)J recombination relies on the O-GlcNAc status of stage-specific proteins during early B-cell development and identified O-GlcNAc-dependent mechanisms as new regulatory components for the development of a diverse antibody repertoire.展开更多
基金supported by a National Research Foundation(NRF)grant from Korea funded by the Ministry of Education(2021R1I1A2057945 to S-KP)the Ministry of Science&ICT(2020R1I1A3073845 to AKP,and 2021R1A2C1012477 to JL)+1 种基金the National Research Council of Science&Technology(NST)(GTL24021-000 to T-DK,GTL24021-400 to HH,and ZYM9382312 to Y-JK as the Postdoctoral Fellowship Program for Young Scientists)the Korea Research Institute of Bioscience and Biotechnology(KRIBB)Research Initiative Program(KGM9942421).
文摘V(D)J recombination secures the production of functional immunoglobulin(Ig)genes and antibody diversity during the early stages of B-cell development through long-distance interactions mediated by cis-regulatory elements and trans-acting factors.O-GlcNAcylation is a dynamic and reversible posttranslational modification of nuclear and cytoplasmic proteins that regulates various protein functions,including DNA-binding affinity and protein-protein interactions.However,the effects of O-GlcNAcylation on proteins involved in V(D)J recombination remain largely unknown.To elucidate this relationship,we downregulated O-GlcNAcylation in a mouse model by administering an O-GlcNAc inhibitor or restricting the consumption of a regular diet.Interestingly,the inhibition of O-GlcNAcylation in mice severely impaired Ig heavy-chain(IgH)gene rearrangement.We identified several factors crucial for V(D)J recombination,including YY1,CTCF,SMC1,and SMC3,as direct targets of O-GlcNAc modification.Importantly,O-GlcNAcylation regulates the physical interaction between SMC1 and SMC3 and the DNA-binding patterns of YY1 at the IgH gene locus.Moreover,O-GlcNAc inhibition downregulated DDX5 protein expression,affecting the functional association of CTCF with its DNA-binding sites at the IgH locus.Our results showed that locus contraction and long-range interactions throughout the IgH locus are disrupted in a manner dependent on the cellular O-GlcNAc level.In this study,we established that V(D)J recombination relies on the O-GlcNAc status of stage-specific proteins during early B-cell development and identified O-GlcNAc-dependent mechanisms as new regulatory components for the development of a diverse antibody repertoire.
