Evidence of the Involvement of O-GlcNAc-modified Human RNA Polymerase II CTD in Transcription in Vitro and in Vivo

Ranuncolo, Stella Maris; Ghosh, Salil; Hanover, John A.; Hart, Gerald W.; Lewis, Brian A.

doi:10.1074/jbc.m111.330910

Cited by 142 publications

(176 citation statements)

References 55 publications

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“…7A, the OGT inhibitor suppressed nascent RNA synthesis in T cells cultured on anti-CD3/CD28-coated coverslips, as measured by metabolic incorporation of a uridine analog. Although this result is consistent with the notion that O-GlcNAc directly affects the activity of RNA polymerase, transcription factors, or epigenetic regulators (63,64), the observed decrease in labeling may also arise indirectly from a defect in T cell activation, which normally leads to increased RNA synthesis (65). Thus, we repeated this experiment using flow cytometry to enable the analysis of resting T cells.…”

Section: Inhibition Of Ogt Blocks Rna Synthesissupporting

confidence: 85%

Global Analysis of O-GlcNAc Glycoproteins in Activated Human T Cells

Lund

Elias

Davis

2016

The Journal of Immunology

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T cell activation in response to Ag is largely regulated by protein posttranslational modifications. Although phosphorylation has been extensively characterized in T cells, much less is known about the glycosylation of serine/threonine residues by O-linked N-acetylglucosamine (O-GlcNAc). Given that O-GlcNAc appears to regulate cell signaling pathways and protein activity similarly to phosphorylation, we performed a comprehensive analysis of O-GlcNAc during T cell activation to address the functional importance of this modification and to identify the modified proteins. Activation of T cells through the TCR resulted in a global elevation of O-GlcNAc levels and in the absence of O-GlcNAc, IL-2 production and proliferation were compromised. T cell activation also led to changes in the relative expression of O-GlcNAc transferase (OGT) isoforms and accumulation of OGT at the immunological synapse of murine T cells. Using a glycoproteomics approach, we identified >200 O-GlcNAc proteins in human T cells. Many of the identified proteins had a functional relationship to RNA metabolism, and consistent with a connection between O-GlcNAc and RNA, inhibition of OGT impaired nascent RNA synthesis upon T cell activation. Overall, our studies provide a global analysis of O-GlcNAc dynamics during T cell activation and the first characterization, to our knowledge, of the O-GlcNAc glycoproteome in human T cells.

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Section: Inhibition Of Ogt Blocks Rna Synthesissupporting

confidence: 85%

Global Analysis of O-GlcNAc Glycoproteins in Activated Human T Cells

Lund

Elias

Davis

2016

The Journal of Immunology

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“…Both OGT and OGA are important in modulating gene transcription (23). For example, RNA polymerase II is modified by O-GlcNAc (24), and O-GlcNAcylation of RNA polymerase II promotes the formation of the transcriptional preinitiation complex (25); however, for transcriptional elongation to proceed OGA needs to remove the O-GlcNAc to facilitate the phosphorylation of the RNA polymerase II C-terminal tail and elongation (25). O-GlcNAcylation is also important for proper protein translation with alterations to cellular O-GlcNAc levels impinging on the function of the translational machinery (26).…”

Section: Discussionmentioning

confidence: 99%

Altering O-Linked β-N-Acetylglucosamine Cycling Disrupts Mitochondrial Function

Tan¹,

Villar²,

Lezi³

et al. 2014

Journal of Biological Chemistry

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“…CTD O-GlcNAcylation was originally found on calf thymus RNA pol II and more recently on human pol II (19,20). Edman degradation of calf thymus pol II indicated that both Thr-4 and Ser-5 of the CTD are O-GlcNAcylated (19), whereas serine to alanine substitutions (20) and mass spectroscopy indicated that Ser-5 was O-GlcNAcylated.…”

Section: Minireview: O-glcnac and Epigeneticsmentioning

confidence: 99%

“…it is a pre-initiation species or a post-elongation species). Focus centered on pol II␥ acting before initiation after finding that OGT and OGA catalytic activity are necessary for transcription in vitro and that inhibition of these enzymes blocks transcription during pre-initiation complex assembly (20). OGT can be detected at promoters and immunoprecipitated with pol II (20).…”

Section: Minireview: O-glcnac and Epigeneticsmentioning

confidence: 99%

“…In contrast, pol IIO is thought to be heavily phosphorylated. A third distinct species of pol II is defined by the O-GlcNAcylation of the CTD (pol II␥) (19,20). pol II␥ was found within the pol IIA population, and in fact, because of the mixing of pol IIA and pol II␥, it has never been clear whether pol IIA or pol II␥ is the initiationspecific species of pol II.…”

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confidence: 99%

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O-GlcNAc and the Epigenetic Regulation of Gene Expression

Lewis¹,

Hanover

2014

Journal of Biological Chemistry

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127

116

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O-GlcNAcylation is an abundant nutrient-driven modification linked to cellular signaling and regulation of gene expression. Utilizing precursors derived from metabolic flux, O-GlcNAc functions as a homeostatic regulator. The enzymes of O-GlcNAc cycling, OGT and O-GlcNAcase, act in mitochondria, the cytoplasm, and the nucleus in association with epigenetic "writers" and "erasers" of the histone code. Both O-GlcNAc and O-phosphate modify repeats within the RNA polymerase II C-terminal domain (CTD). By communicating with the histone and CTD codes, O-GlcNAc cycling provides a link between cellular metabolic status and the epigenetic machinery. Thus, O-GlcNAcylation is poised to influence transgenerational epigenetic inheritance.Intermediary metabolism, the process by which nutrients are converted into cellular biomass, is an interwoven network of biochemical reactions allowing reproduction, development, and response to the environment. The intermediary metabolic network is highly conserved and includes every cellular process ranging from DNA replication to transcription and translation to enzyme regulation. Epigenetics, the study of how genes may alter phenotypes beyond their ability to genetically encode information, is ultimately linked to intermediary metabolism (1). Many enzymes that participate in epigenetic gene regulation depend upon co-substrates produced by cellular metabolism, thus providing a potential link between metabolism and gene regulation (2). Mitochondria, key players in these metabolic inter-conversions, exhibit a pattern of cytoplasmic inheritance distinct from Mendelian inheritance of genes encoded in the nucleus (3). O-GlcNAcylation is a key integrator of cellular nutritional status and occurs in the nucleus, cytoplasm, and mitochondrion. O-GlcNAc transferase (OGT) 3 utilizes UDP-GlcNAc to catalyze the addition of O-GlcNAc to target proteins. UDP-GlcNAc is the end product of the hexosamine biosynthetic pathway (HSP), a series of enzymatic reactions requiring key metabolites, including glucose, glutamine, ATP, and acetyl-CoA (4). The nutrient-derived precursors render the synthesis of UDP-GlcNAc and subsequent O-GlcNAc addition by OGT nutrient-responsive. The O-GlcNAcase (OGA; MGEA5) removes the O-GlcNAc modification, and evidence suggests that its transcription and activity is highly regulated. Both enzymes of O-GlcNAc cycling contain domains that allow them to bind to epigenetic modifiers (5, 6). As illustrated in Fig. 1, UDP-GlcNAc is central both to the formation of O-GlcNAc but also to the synthesis of membrane and secretory glycoproteins that perform essential roles in extracellular signaling. The intracellular O-GlcNAc modification plays a role in signaling, leading to growth and apoptosis (7-9), metabolism (10, 11), and the cell cycle (12)(13)(14). In addition, O-GlcNAc has been implicated in translation (15), circadian rhythm (16), the establishment of molecular memory in neurons (17), and calmodulinkinase signaling (16). The focus of this review is the role of O-GlcNAc in transcripti...

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Evidence of the Involvement of O-GlcNAc-modified Human RNA Polymerase II CTD in Transcription in Vitro and in Vivo

Cited by 142 publications

References 55 publications

Global Analysis of O-GlcNAc Glycoproteins in Activated Human T Cells

Global Analysis of O-GlcNAc Glycoproteins in Activated Human T Cells

Altering O-Linked β-N-Acetylglucosamine Cycling Disrupts Mitochondrial Function

O-GlcNAc and the Epigenetic Regulation of Gene Expression

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