Quantitative and Site-Specific Chemoproteomic Profiling of Protein O‑GlcNAcylation in the Cell Cycle

Mammalian
cell cycle is a central process for tissue growth and
maintenance. Protein O-linked β-N-acetylglucosamine
(O-GlcNAc) modification has been found to occur on several important
cell cycle regulators. However, the O-GlcNAcylated proteome has not
been extensively profiled during cell cycle progression. Herein, we
report a quantitative profiling of protein O-GlcNAcylation sites in
cell proliferation, by using an O-GlcNAc chemoproteomic strategy.
In HeLa cells, a total of 902, 439, and 872 high-confidence O-GlcNAcylation
sites distributed on 414, 265, and 425 proteins are identified in
the interphase, early mitosis, and mitotic exit stages, respectively.
The identified O-GlcNAcylation events occur on a variety of important
regulators, which are involved in the processes of cell division,
DNA repair, and cell death. Furthermore, we show that O-GlcNAcylation
is dynamically regulated in a cell cycle stage-dependent manner. Our
results provide a valuable resource for investigating the functional
roles of O-GlcNAc in the mammalian cell cycle.