O-GlcNAcylation regulates the DNA damage repair function of MND1 in breast cancer.

INTRODUCTION

As a meiosis-specific gene, Meiotic Nuclear Divisions 1 (MND1) plays a crucial role in homologous recombination and is generally silenced in somatic cells. The expression pattern and functional significance of MND1 in breast cancer (BRCA) have not been elucidated. This study systematically investigated the expression, subcellular localization, and pathological function of MND1 in BRCA, with particular focus on the post-translational regulation of MND1 through O-GlcNAcylation (O-GlcNAc) and its underlying molecular mechanisms.

METHODS

Immunohistochemical analysis and subcellular fractionation assays were performed to quantify MND1 expression levels and determine its intracellular localization in BRCA tissues and matched adjacent normal tissues. A drug-induced double-strand break (DSB) model combined with pH2A.X quantification were used to establish the functional correlation between MND1 and DSB repair efficiency. An in situ transplantation model of BRCA was established to evaluate the tumorigenic potential of MND1. O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) inhibitors, supplemented with site-directed mutagenesis (T121A mutation), were implemented to dissect the regulatory effects of O-GlcNAc on MND1 protein dynamics and functional competence.

RESULTS

MND1 was aberrantly overexpressed and predominantly localized in the nucleus in BRCA cells. The DSB levels induced by DOX in BRCA cells were negatively correlated with the protein levels of MND1. knockdown of MND1 significantly inhibited tumor growth in mice, elevated the DSB levels, and reduced Ki67 levels in tumors. Immunoprecipitation coupled with in vitro glycosylation assays confirmed site-specific O-GlcNAc modification at MND1-T121. Pharmacological inhibition of OGT and T121A mutagenesis dramatically reduced MND1 protein stability and impaired nuclear localization. T121A mutation compromised MND1-mediated DSB repair capacity, resulting in persistently elevated pH2A.X levels. Inhibition of O-GlcNAc on MND1-T121 augmented MND1-HOP2 interaction, which led to a significant structural perturbation between the MND1-HOP2 complex.

CONCLUSIONS

Our findings elucidate nuclear MND1 as a BRCA-specific HR activator stabilized by O-GlcNAc-mediated ubiquitination resistance. Thr121 O-GlcNAcylation facilitates MND1 nuclear retention and modulates HOP2 interaction dynamics, critically enabling HR-mediated repair. Loss of this modification disrupts repair fidelity, exposing a targetable vulnerability in BRCA. Future studies should characterize the structural basis of O-GlcNAc-dependent MND1-HOP2 regulation and its therapeutic implications.