Nuclear localization of tetrahydrobiopterin biosynthetic enzymes

Elzaouk, Lina; Laufs, Stephanie; Heerklotz, Dirk; Leimbacher, Walter; Blau, Nenad; Résibois, A.; Thöny, Beat

doi:10.1016/j.bbagen.2003.10.015

Cited by 26 publications

(17 citation statements)

References 40 publications

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“…NO orchestrated the induction of the iNOS gene through an ordered cascade of biochemical events. In accordance with the increasing armamentarium of serpentine receptor downstream effectors (see above) along with co-factors and proteins regulating eNOS activity (Ca 2ϩ ions, BH 4 -biosynthetic enzymes, Akt, calmodulin) (16,38,39) that are found in the nucleus, our results indicate that in the nuclear compartment eNOS-derived NO stimulates a sGC/cGMP/ PKG pathway to induce nuclear calcium transients, consistent with activation of calcium channels by cGMP/PKG in hepatocytes (40,41). This action in turn initiates MAPK (p42/44)-sensitive NF-B activation and finally induces iNOS gene expression (Figs.…”

Section: Discussionmentioning

confidence: 99%

Nitric Oxide Signaling via Nuclearized Endothelial Nitric-oxide Synthase Modulates Expression of the Immediate Early Genes iNOS and mPGES-1

Gobeil

Zhu

Brault

et al. 2006

Journal of Biological Chemistry

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Stimulation of freshly isolated rat hepatocytes with lysophosphatidic acid (LPA) resulted in LPA 1 receptor-mediated and nitricoxide-dependent up-regulation of the immediate early genes iNOS (inducible nitric-oxide synthase (NOS)) and mPGES-1 (microsomal prostaglandin E synthase-1). Because LPA is a ligand for both cell surface and intracellular receptor sites and a potent endothelial NOS (eNOS) activator, we hypothesized that NO derived from activated nuclearized eNOS might participate in gene regulation. Herein we show, by confocal microscopy performed on porcine cerebral endothelial cells expressing native LPA 1 -receptor and eNOS and on HTC4 rat hepatoma cells co-transfected with recombinant human LPA 1 -receptor and fused eNOS-GFP cDNA, a dynamic eNOS translocation from peripheral to nuclear regions upon stimulation with LPA. Nuclear localization of eNOS and its downstream effector, soluble guanylate cyclase, were demonstrated in situ in rat liver specimens by immunogold labeling using specific antibodies. Stimulation of this nuclear fraction with LPA and the NO donor sodium nitroprusside resulted, respectively, in increased production of nitrite (and eNOS phosphorylation) and cGMP; these separate responses were also correspondingly blocked by NOS inhibitor L-NAME and soluble guanylate cyclase inhibitor ODQ. In addition, sodium nitroprusside evoked a sequential increase in nuclear Ca 2؉ transients, activation of p42 MAPK, NF-B binding to DNA consensus sequence, and dependent iNOS RNA. This study describes a hitherto unrecognized molecular mechanism by which nuclear eNOS through ensuing NO modulates nuclear calcium homeostasis involved in gene transcription-associated events. Moreover, our findings strongly support the concept of the nucleus as an autonomous signaling compartment.Nitric oxide (NO) is a short-lived uncharged free radical involved in numerous complex biological processes such as blood pressure regulation, vascular inflammation, cell-mediated cytotoxicity, and survival (1, 2). One of the most biologically relevant actions of NO is its binding to the heme moiety in the heterodimeric enzyme soluble guanylate cyclase (sGC), 3 which leads to the production of the intracellular second messenger molecule cGMP and activation of cGMP-dependent protein kinases G (PKG). However, NO can also interact with and modify the bioactivity of a number of protein macromolecules through a series of reversible and nonreversible chemical reactions (e.g. S-, N-, and hemenitrosylation, tyrosine and tryptophan nitration) providing multifaceted regulatory mechanisms for cellular functions (3,4). NO is synthesized from L-arginine by a tripartite family of nitric-oxide synthase (NOS) isozymes composed of NOS-1 (nNOS), NOS-2 (iNOS), and NOS-3 (eNOS) originally found in brain, macrophages, and endothelium, respectively, and later discovered, for each NOS, in many other cell types (5).NOS isozymes, especially eNOS, are highly regulated by a number of mechanisms. Emerging findings show that eNOS bioactivity and mobilization to...

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Section: Discussionmentioning

confidence: 99%

Nitric Oxide Signaling via Nuclearized Endothelial Nitric-oxide Synthase Modulates Expression of the Immediate Early Genes iNOS and mPGES-1

Gobeil

Zhu

Brault

et al. 2006

Journal of Biological Chemistry

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“…5A). Using the specific antibody SZ28 against PTPS protein [Elzaouk et al, 2004], Figure 2. Result of the ex vivo splicing in the minigene assay using constructs with wild-type sequences, the mutation c.164À712A4T from patient MD355, and the changes c.164À714C4A and c.164À715T4A.…”

Section: Reversion Of the Splicing Defect And Restoration Of Ptps Enzmentioning

confidence: 99%

“…Poinceau staining was used to monitor equal loading of protein. Immunodetection was carried out using antihuman PTPS polyclonal antibody SZ28 (1:500 diluted) [Elzaouk et al, 2004] followed by a second antibody (conjugated goat antirabbit IgG horseradish peroxidase, diluted 1:10,000; Santa Cruz Biotechnology Inc., Santa Cruz, CA). The detection was made using the Enhanced Chemiluminescence System (GE Healthcare, Buckinghamshire, UK).…”

Section: Ptps Activity Pterin Quantification and Protein Immunodetementioning

confidence: 99%

Pseudoexon exclusion by antisense therapy in 6-pyruvoyl-tetrahydropterin synthase deficiency

et al. 2011

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Antisense oligonucleotide therapy to modulate splicing mutations in inherited diseases is emerging as a treatment option also for metabolic defects. In this article, we report the effect of cellular antisense therapy to suppress pseudoexon activation in primary dermal fibroblasts from patients with mutations in the PTS gene encoding 6-pyruvoyltetrahydropterin synthase (PTPS), which leads to tetrahydrobiopterin and monoamine neurotransmitter deficiency. Pathogenic inclusion of SINE or LINE-derived cryptic exons in different PTPS patients due to the intronic mutations c.84-322A>T, c.163 + 695_163 + 751del57, or c.164-712A>T was demonstrated by transcript analysis in fibroblasts and minigene ex vivo assays. Antisense morpholino oligonucleotides (AMOs) directed to the pseudoexons 3' or 5' splice sites were designed with the aim of preventing the pathological pseudoexon inclusion. At the time of AMO transfection, we investigated patients' cells for correct PTS-mRNA splicing and functional recovery of the PTPS protein. Transcriptional profiling after 24 hr posttransfection revealed a dose- and sequence-specific recovery of normal splicing. Furthermore, PTPS enzyme activity in all three patients' fibroblasts and the pterin profile were close to normal values after antisense treatment. Our results demonstrate proof-of-concept for pseudoexon exclusion therapy using AMO in inherited metabolic disease.

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“…The three NOS isoforms have all been observed in cell nuclei under pathophysiological conditions [99–103]. Guanylyl cyclase [104], subunits of the sGC [105], cGMP production [105], calmodulin [106] and tetrahydrobiopterin biosynthetic enzymes [107] have also been detected in nuclei, suggesting that nuclear production of nitric oxide and the subsequent activation of its downstream targets is possible.…”

Section: Translocation Of Nitric Oxide Synthases Among Cellular Compamentioning

confidence: 99%

Subcellular and cellular locations of nitric oxide synthase isoforms as determinants of health and disease

Villanueva

Giulivi

2010

Free Radical Biology and Medicine

192

127

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The effects of nitric oxide in biological systems depend on its steady-state concentration and where it is being produced. The organ where nitric oxide is produced is relevant, and within the organ, which types of cells are actually contributing to this production seem to play a major determinant of its effect. Subcellular compartmentalization of specific nitric-oxide synthase enzymes has been shown to play a major role in health and disease. Pathophysiological conditions affect the cellular expression and localization of nitric oxide synthases, which in turn alter organ cross talk. In this study, we described the compartmentalization of nitric oxide in organs, cells and subcellular organelles, and how its localization relates to several relevant clinical conditions. Understanding the complexity of the compartmentalization of nitric oxide production and the implications of this compartmentalization in terms of cellular targets and downstream effects will eventually contribute toward the development of better strategies for treating or preventing pathological events associated with the increase, inhibition or mislocalization of nitric oxide production.

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Nuclear localization of tetrahydrobiopterin biosynthetic enzymes

Cited by 26 publications

References 40 publications

Nitric Oxide Signaling via Nuclearized Endothelial Nitric-oxide Synthase Modulates Expression of the Immediate Early Genes iNOS and mPGES-1

Nitric Oxide Signaling via Nuclearized Endothelial Nitric-oxide Synthase Modulates Expression of the Immediate Early Genes iNOS and mPGES-1

Pseudoexon exclusion by antisense therapy in 6-pyruvoyl-tetrahydropterin synthase deficiency

Subcellular and cellular locations of nitric oxide synthase isoforms as determinants of health and disease

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