Dynamic Glycosylation Governs the Vertebrate COPII Protein Trafficking Pathway

The COPII coat complex, which mediates secretory cargo trafficking from the endoplasmic reticulum, is a key control point for subcellular protein targeting. Because misdirected proteins cannot function, protein sorting by COPII is critical for establishing and maintaining normal cell and tissue homeostasis. Indeed, mutations in COPII genes cause a range of human pathologies, including cranio-lenticulo-sutural dysplasia (CLSD), which is characterized by collagen trafficking defects, craniofacial abnormalities, and skeletal dysmorphology. Detailed knowledge of the COPII pathway is required to understand its role in normal cell physiology and to devise new treatments for disorders in which it is disrupted. However, little is known about how vertebrates dynamically regulate COPII activity in response to developmental, metabolic, or pathological cues. Several COPII proteins are modified by O-linked β-<i>N</i>-acetylglucosamine (O-GlcNAc), a dynamic form of intracellular protein glycosylation, but the biochemical and functional effects of these modifications remain unclear. Here, we use a combination of chemical, biochemical, cellular, and genetic approaches to demonstrate that site-specific O-GlcNAcylation of COPII proteins mediates their protein–protein interactions and modulates cargo secretion. In particular, we show that individual O-GlcNAcylation sites of SEC23A, an essential COPII component, are required for its function in human cells and vertebrate development, because mutation of these sites impairs SEC23A-dependent <i>in vivo</i> collagen trafficking and skeletogenesis in a zebrafish model of CLSD. Our results indicate that O-GlcNAc is a conserved and critical regulatory modification in the vertebrate COPII-dependent trafficking pathway

Microbiota-dependent histone butyrylation in the mammalian intestine

This dataset includes supporting data for figure generation in the associated manuscript. The dataset contents include images and excel files. The image files were generated from immunoblotting (related to Figs. 1B, 2, 3E, and Extended Data Fig. 2) and immunofluorescence experiments (related to Fig. 1E). Each immunoblotting image has two files associated with each figure panel: the file name lists the figure, primary antibody, and either "ECL" = chemiluminescence only, or "full image" = chemiluminescence plus color image of entire gel, which may also include notations (such as protein ladder position or other information to orient the viewer). The excel files include quantification for figures (related to Figs. 2, 3D, Extended Data Fig. 3A), summaries of 16S sequencing results (related to Extended Data Fig. 3B), gene ontology analysis of RNA-seq data (related to Extended Data Fig. 4E) and ChIP-seq data (related to Extended Data Fig. 6), and summaries of metabolomics mass spectrometry data (related to Extended Data Fig. 5)