Hexosamine biosynthetic pathway and O-GlcNAc-processing enzymes regulate daily rhythms in protein O-GlcNAcylation

Xianhui Liu,Christine A Tabuloc,Thu M Pham,Joanna C Chiu,Ying H Li,Ivana Blaženović,Jian Ji,Adam J Contreras,Oliver Fiehn

doi:10.1038/s41467-021-24301-7

Abstract

The integration of circadian and metabolic signals is essential for maintaining robust circadian rhythms and ensuring efficient metabolism and energy use. Using Drosophila as an animal model, we show that cellular protein O-GlcNAcylation exhibits robust 24-hour rhythm and represents a key post-translational mechanism that regulates circadian physiology. We observe strong correlation between protein O-GlcNAcylation rhythms and clock-controlled feeding-fasting cycles, suggesting that O-GlcNAcylation rhythms are primarily driven by nutrient input. Interestingly, daily O-GlcNAcylation rhythms are severely dampened when we subject flies to time-restricted feeding at unnatural feeding time. This suggests the presence of clock-regulated buffering mechanisms that prevent excessive O-GlcNAcylation at non-optimal times of the day-night cycle. We show that this buffering mechanism is mediated by the expression and activity of GFAT, OGT, and OGA, which are regulated through integration of circadian and metabolic signals. Finally, we generate a mathematical model to describe the key factors that regulate daily O-GlcNAcylation rhythm.

Highlights

The integration of circadian and metabolic signals is essential for maintaining robust circadian rhythms and ensuring efficient metabolism and energy use
Using chemoenzymatic labeling[27], a strategy we have previously employed to examine PERIOD (PER) O-GlcNAcylation[26], we conjugated biotin tags to OGlcNAc groups on nuclear proteins extracted from wild type (WT; w1118) male fly bodies
Our results showed that O-GlcNAcylation of nuclear proteins exhibited a robust daily rhythm, suggesting that timely metabolic input may regulate the function of cellular proteins through this post-translational modification

Summary

Introduction

The integration of circadian and metabolic signals is essential for maintaining robust circadian rhythms and ensuring efficient metabolism and energy use. The circadian clock has been shown to drive daily rhythmic expression of metabolic genes involved in glycolysis, pentose phosphate pathway, gluconeogenesis, lipid oxidation and storage[1,7] Once metabolized, macronutrients such as sugars, amino acids and lipids, or the lack of nutrients during fasting period can in turn regulate appropriate nutrient-sensing signaling pathways, such as those regulated by insulin/target of rapamycin (TOR), adenosine-monophosphate-activated protein kinase (AMPK), glucagon, adipokines, and autophagy, to orchestrate downstream physiological functions[1,6,8]. Together with the fact that there is extensive interplay between O-GlcNAcylation and phosphorylation to regulate cellular protein functions[16,17] and recent findings that a significant portion of the phosphoproteomes in mice and flies exhibit daily oscillations[20,21,22], we sought to investigate if protein O-GlcNAcylation represents an important post-translational mechanism that integrates circadian and metabolic regulation to coordinate circadian physiology

Methods

Results

Conclusion