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Dataset on the formation of Thioredoxin interacting protein (Txnip) containing redox sensitive high molecular weight nucleoprotein complexes.

ABSTRACT: This dataset is supplementary to the submitted research by Ref. [1]. RNAs were extracted from high molecular weight complexes, prepared with 100 kDa filtration of HEK293 Tet-on cells stably transfected with either F-HA-Txnip-V5-His or control vector. Cells were stimulated with 1 ?g/mL doxycycline for 24 h, followed by overnight stimulation with 100 ?M 4-thiouridine (4sU), 20 mM glucose, and 1 ?M bortezomib for 14h. The extracted RNAs from Txnip overexpressing cells compared with control cells was analyzed by RNA-seq. Differentially expressed mRNAs, long noncoding RNAs (lncRNA) and transcripts of uncertain coding potential (TUCPs) are shown. Gene ontology and KEGG enrichment of these differential expressed RNAs is presented.

SUBMITTER: Hirata CL

PROVIDER: S-EPMC6926129 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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Dataset on the formation of Thioredoxin interacting protein (Txnip) containing redox sensitive high molecular weight nucleoprotein complexes.

Hirata Cristiane Lumi CL Ito Shinji S Masutani Hiroshi H

Data in brief 20191129

This dataset is supplementary to the submitted research by Ref. [1]. RNAs were extracted from high molecular weight complexes, prepared with 100 kDa filtration of HEK293 Tet-on cells stably transfected with either F-HA-Txnip-V5-His or control vector. Cells were stimulated with 1 μg/mL doxycycline for 24 h, followed by overnight stimulation with 100 μM 4-thiouridine (4sU), 20 mM glucose, and 1 μM bortezomib for 14h. The extracted RNAs from Txnip overexpressing cells compared with control cells wa ...[more]

PMID: 31890777

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Project description:Thioredoxin-interacting protein (TXNIP) regulates critical biological processes including inflammation, stress and apoptosis. TXNIP is upregulated by glucose and is a critical mediator of hyperglycemia-induced beta-cell apoptosis in diabetes. In contrast, the saturated long-chain fatty acid palmitate, although toxic to the beta-cell, inhibits TXNIP expression. The mechanisms involved in the opposing effects of glucose and fatty acids on TXNIP expression are unknown. We found that both palmitate and oleate inhibited TXNIP in a rat beta-cell line and islets. Palmitate inhibition of TXNIP was independent of fatty acid beta-oxidation or esterification. AMP-activated protein kinase (AMPK) has an important role in cellular energy sensing and control of metabolic homeostasis; therefore we investigated its involvement in nutrient regulation of TXNIP. As expected, glucose inhibited whereas palmitate stimulated AMPK. Pharmacologic activators of AMPK mimicked fatty acids by inhibiting TXNIP. AMPK knockdown increased TXNIP expression in presence of high glucose with and without palmitate, indicating that nutrient (glucose and fatty acids) effects on TXNIP are mediated in part via modulation of AMPK activity. TXNIP is transcriptionally regulated by carbohydrate response element-binding protein (ChREBP). Palmitate inhibited glucose-stimulated ChREBP nuclear entry and recruitment to the Txnip promoter, thereby inhibiting Txnip transcription. We conclude that AMPK is an important regulator of Txnip transcription via modulation of ChREBP activity. The divergent effects of glucose and fatty acids on TXNIP expression result in part from their opposing effects on AMPK activity. In light of the important role of TXNIP in beta-cell apoptosis, its inhibition by fatty acids can be regarded as an adaptive/protective response to glucolipotoxicity. The finding that AMPK mediates nutrient regulation of TXNIP may have important implications for the pathophysiology and treatment of diabetes.

Project description:A classical hallmark of Parkinson's disease (PD) pathogenesis is the accumulation of misfolded alpha-synuclein (αSyn) within Lewy bodies and Lewy neurites, although its role in microglial dysfunction and resultant dopaminergic (DAergic) neurotoxicity is still elusive. Previously, we identified that protein kinase C delta (PKCδ) is activated in post mortem PD brains and experimental Parkinsonism and that it participates in reactive microgliosis; however, the relationship between PKCδ activation, endoplasmic reticulum stress (ERS) and the reactive microglial activation state in the context of α-synucleinopathy is largely unknown. Herein, we show that oxidative stress, mitochondrial dysfunction, NLR family pyrin domain containing 3 (NLRP3) inflammasome activation, and PKCδ activation increased concomitantly with ERS markers, including the activating transcription factor 4 (ATF-4), serine/threonine-protein kinase/endoribonuclease inositol-requiring enzyme 1α (p-IRE1α), p-eukaryotic initiation factor 2 (eIF2α) as well as increased generation of neurotoxic cytokines, including IL-1β in aggregated αSynagg-stimulated primary microglia. Importantly, in mouse primary microglia-treated with αSynagg we observed increased expression of Thioredoxin-interacting protein (TXNIP), an endogenous inhibitor of the thioredoxin (Trx) pathway, a major antioxidant protein system. Additionally, αSynagg promoted interaction between NLRP3 and TXNIP in these cells. In vitro knockdown of PKCδ using siRNA reduced ERS and led to reduced expression of TXNIP and the NLRP3 activation response in αSynagg-stimulated mouse microglial cells (MMCs). Additionally, attenuation of mitochondrial reactive oxygen species (mitoROS) via mito-apocynin and amelioration of ERS via the eIF2α inhibitor salubrinal (SAL) reduced the induction of the ERS/TXNIP/NLRP3 signaling axis, suggesting that mitochondrial dysfunction and ERS may act in concert to promote the αSynagg-induced microglial activation response. Likewise, knockdown of TXNIP by siRNA attenuated the αSynagg-induced NLRP3 inflammasome activation response. Finally, unilateral injection of αSyn preformed fibrils (αSynPFF) into the striatum of wild-type mice induced a significant increase in the expression of nigral p-PKCδ, ERS markers, and upregulation of the TXNIP/NLRP3 inflammasome signaling axis prior to delayed loss of TH+ neurons. Together, our results suggest that inhibition of ERS and its downstream signaling mediators TXNIP and NLRP3 might represent novel therapeutic avenues for ameliorating microglia-mediated neuroinflammation in PD and other synucleinopathies.

Dataset Information

Dataset on the formation of Thioredoxin interacting protein (Txnip) containing redox sensitive high molecular weight nucleoprotein complexes.

Publications

Dataset on the formation of Thioredoxin interacting protein (Txnip) containing redox sensitive high molecular weight nucleoprotein complexes.

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