Profiling of Protein <i>O</i>‑GlcNAcylation in Murine CD8⁺ Effector- and Memory-like T Cells

During an acute infection, antigenic stimulation leads to activation, expansion, and differentiation of naïve CD8⁺ T cells, first into cytotoxic effector cells and eventually into long-lived memory cells. T cell antigen receptors (TCRs) detect antigens on antigen-presenting cells (APCs) in the form of antigenic peptides bound to major histocompatibility complex I (MHC-I)-encoded molecules and initiate TCR signal transduction network. This process is mediated by phosphorylation of many intracellular signaling proteins. Protein <i>O</i>-GlcNAc modification is another post-translational modification involved in this process, which often has either reciprocal or synergistic roles with phosphorylation. In this study, using a chemoenzymatic glycan labeling technique and proteomics analysis, we compared protein <i>O</i>-GlcNAcylation of murine effector and memory-like CD8⁺ T cells differentiated <i>in vitro</i>. By quantitative proteomics analysis, we identified 445 proteins that are significantly regulated in either effector- or memory-like T cell subsets. Furthermore, qualitative and quantitative analysis identified highly regulated protein clusters that suggest involvement of this post-translational modification in specific cellular processes. In effector-like T cells, protein <i>O</i>-GlcNAcylation is heavily involved in transcriptional and translational processes that drive fast effector T cells proliferation. During the formation of memory-like T cells, protein <i>O</i>-GlcNAcylation is involved in a more specific, perhaps more targeted regulation of transcription, mRNA processing, and translation. Significantly, <i>O</i>-GlcNAc plays a critical role as part of the “histone code” in both CD8⁺ T cells subgroups

Profiling of Protein <i>O</i>‑GlcNAcylation in Murine CD8⁺ Effector- and Memory-like T Cells

During an acute infection, antigenic stimulation leads to activation, expansion, and differentiation of naïve CD8⁺ T cells, first into cytotoxic effector cells and eventually into long-lived memory cells. T cell antigen receptors (TCRs) detect antigens on antigen-presenting cells (APCs) in the form of antigenic peptides bound to major histocompatibility complex I (MHC-I)-encoded molecules and initiate TCR signal transduction network. This process is mediated by phosphorylation of many intracellular signaling proteins. Protein <i>O</i>-GlcNAc modification is another post-translational modification involved in this process, which often has either reciprocal or synergistic roles with phosphorylation. In this study, using a chemoenzymatic glycan labeling technique and proteomics analysis, we compared protein <i>O</i>-GlcNAcylation of murine effector and memory-like CD8⁺ T cells differentiated <i>in vitro</i>. By quantitative proteomics analysis, we identified 445 proteins that are significantly regulated in either effector- or memory-like T cell subsets. Furthermore, qualitative and quantitative analysis identified highly regulated protein clusters that suggest involvement of this post-translational modification in specific cellular processes. In effector-like T cells, protein <i>O</i>-GlcNAcylation is heavily involved in transcriptional and translational processes that drive fast effector T cells proliferation. During the formation of memory-like T cells, protein <i>O</i>-GlcNAcylation is involved in a more specific, perhaps more targeted regulation of transcription, mRNA processing, and translation. Significantly, <i>O</i>-GlcNAc plays a critical role as part of the “histone code” in both CD8⁺ T cells subgroups