Published online before print November 1, 2007, doi: 10.1161/CIRCRESAHA.107.157305
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© 2008 American Heart Association, Inc.
Molecular Medicine |
Nitric Oxide Modulates Chromatin Folding in Human Endothelial Cells via Protein Phosphatase 2A Activation and Class II Histone Deacetylases Nuclear Shuttling
From the Laboratorio di Biologia Vascolare e Terapia Genica (B.I., F.S.), Centro Cardiologico Fondazione "I. Monzino", IRCCS, Milan; Istituto di Ricerche di Biologia Molecolare I.R.B.M. P. Angeletti (C.D.R., C.S., P.G.), Via Pontina km 30 600, Pomezia, Rome; Laboratorio di Patologia Vascolare (C.C., J.R., G.R., F.M., M.C.C.), Istituto Dermopatico dell’ Immacolata-IRCCS, Rome; Università di Siena (M.P.), Siena; Dipartimento di Cardiochirurgia (P.B.), Centro Cardiologico Fondazione "I. Monzino", IRCCS, Milan; Istituto Pasteur-Fondazione Cenci Bolognetti (D.R., S.V., A.M.), Dipartimento Studi Farmaceutici Università degli Studi di Roma "La Sapienza", Rome, Italy.
Correspondence to Barbara Illi, PhD, Centro Cardiologico Fondazione "I. Monzino", Milan, Italy. E-mail b.illi{at}idi.it
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Abstract |
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Nitric oxide (NO) modulates important endothelial cell (EC) functions and gene expression by a molecular mechanism which is still poorly characterized. Here we show that in human umbilical vein ECs (HUVECs) NO inhibited serum-induced histone acetylation and enhanced histone deacetylase (HDAC) activity. By immunofluorescence and Western blot analyses it was found that NO induced class II HDAC4 and 5 nuclear shuttling and that class II HDACs selective inhibitor MC1568 rescued serum-dependent histone acetylation above control level in NO-treated HUVECs. In contrast, class I HDACs inhibitor MS27–275 had no effect, indicating a specific role for class II HDACs in NO-dependent histone deacetylation. In addition, it was found that NO ability to induce HDAC4 and HDAC5 nuclear shuttling involved the activation of the protein phosphatase 2A (PP2A). In fact, HDAC4 nuclear translocation was impaired in ECs expressing small-t antigen and exposed to NO. Finally, in cells engineered to express a HDAC4-Flag fusion protein, NO induced the formation of a macromolecular complex including HDAC4, HDAC3, HDAC5, and an active PP2A. The present results show that NO-dependent PP2A activation plays a key role in class II HDACs nuclear translocation.
Key Words: nitric oxide • endothelial cells • histone deacetylases • chromatin
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Introduction |
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In endothelial cells (ECs) laminar shear stress (SS), the tangential component of hemodynamic forces, enhances NO production via endothelial nitric oxide synthase (eNOS) activation.1 SS-dependent transcriptional responses are partly attributable to NO production,2 and NO has been demonstrated to downregulate gene expression in endothelial cells.3 Nevertheless, the mechanism by which NO regulates gene expression is still unclear.
Changes in chromatin folding are the prerequisite for genes to be turned "on" and "off".4 Histones acetyltransferases (HATs) and deacetylases (HDACs) are histones modifying enzymes involved in the regulation of chromatin unwinding and wrapping, respectively. HAT activity is mainly linked to transcriptional activation, because acetylated histone tails decrease their affinity for DNA, facilitating the recruitment of other chromatin associated transcriptional complexes. HDACs catalyze the removal of acetyl groups from histone tails, compressing chromatin and promoting the repression of transcription.5 The reversible nature of acetylation allows chromatin structure to be tightly regulated to permit the fine tuning of gene expression and a disregulated histone deacetylase activity, together with inappropriate epigenetic patterns, have been recently associated with cancer.6,7 Four classes of histone deacetylases are currently known,5 being class II HDACs mainly involved in the regulation of differentiation programs.8 Although the role of histone deacetylases in the biology of the cardiovascular system is still poorly characterized, different HDACs have been involved in the differentiation of stem cells into endothelial cells9 or in the commitment of endothelial progenitor cells10 and in the maintenance of vascular integrity.11
Our previous work demonstrated that SS regulates gene expression in human ECs and directs embryonic stem cell (ES) differentiation toward the cardiovascular lineage, two processes both associated to the activation of HATs and the opening of chromatin.12,13 In those experiments, SS-dependent histone acetylation and HATs activation was transient, showing a peak between 1 and 2 hours of SS and declining shortly after. In cells treated with the histone deacetylase inhibitor Trichostatin A (TSA), however, histone acetylation remained elevated beyond the 4 hours time point.12 Based on this observation, we hypothesized a potential involvement of a HDAC-dependent activity in this process, in a time frame compatible with NO production.14
In the present study, we show that NO induces class II HDAC4 and 5 nuclear localization in human endothelial cells. This phenomenon is associated with a decrease in histone acetylation and c-fos expression and may provide a novel molecular mechanism for NO-dependent effect on gene expression.3
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Materials and Methods |
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Cell Culture and Treatments
HUVEC were cultured as previously described.12 Treatments were performed as described in supplemental Materials and Methods (available online at http://circres.ahajournals.org).
HEK 293 cells were grown at 37°C in a 5% CO2 atmosphere in complete Dulbecco’s Modified Eagle Medium (GIBCO) containing 0.11g/L Pyridoxine, complemented with 2 mmol/L L-Glutamine, 0.1 mg/mL Penicillin-Streptomycin, 10% (v/v) Fetal Calf Serum (FCS, GIBCO), 250 µg/mL G418, 0.5 µg/mL Puromycin, and 100 µg/mL Hygromycin. Small-t antigen adenoviral and retroviral infection of HUVECs were performed as previously described.15
Immunofluorescence
Immunofluorescences were performed as previously described.13
Cell Extracts and Western Blot
For a detailed protocol of cell fractionation procedure see supplemental Materials and Methods. Western blots were performed as previously described.12,13
HDAC Assay
HDAC assays were performed by using the HDAC activity assay Kit (Upstate Biotechnology) according to the manufacturer’s instructions. For a detailed protocol see supplemental Materials and Methods.
Phosphatase Assay
Phosphatase (PPase) assay were performed by using the Ser/Threo Phosphatase Assay System (Promega) according to the manufacturer’s instructions for the detection of PP2A-specific activity. Total cell extracts were performed by using a standard RIPA buffer, without phosphatase inhibitors.
Plasmid and HDAC4-Flag Purification
HDAC4-Flag was cloned in a pTRE2 hyg vector (Clontech) and stably transfected into an HEK 293 EBNA-1 cell line constitutively expressing both Tet rtTA2s-S2 activator and rTS repressor. To express HDAC4, cells were stimulated 24 hours with 1 µg/mL doxocyclin. After induction, cells were collected and the protein was absorbed onto an anti-Flag resin (Sigma) before competitive elution with a 3x Flag peptide, according to manufacturer instructions.
Confocal Analysis
Confocal analysis, for the detection of HDAC4 and PP2A colocalization, was performed by using an anti-HDAC4 antibody (Santa Cruz Biotechnology) and an anti-PP2Ac antibody (Transduction Laboratory). Nuclei were stained with Topro3 dye. Samples were analyzed using a Zeiss LSM510 Meta Confocal Microscope. Lasers’ power, beam splitters, filter settings, pinhole diameters, and scan mode were the same for all examined samples of each sample. Fields reported in the figure are representative of all examined fields.
Statistical Analysis
Statistical analysis was performed as previously described.12,13
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Results |
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NO Induces Histones Deacetylation in Human Endothelial Cells
SS rapidly induced histone H3 acetylation in human ECs and mouse ES.12,13 The phenomenon was transient, reaching a peak between 1 and 2 hours and decreasing at the 4 hours time point. To investigate whether a link between NO production and histone deacetylation may exist, HUVECs were exposed to SS for 1 to 4 hours in the presence or absence of the nitric oxide synthase inhibitor S-methylthiosourea (SMT). Western blot analysis was performed to detect acetylated histones. SS-dependent histone H3 acetylation on lysine 14 (K14) was detectable at the 1 hour time point, as expected. Surprisingly, SMT enhanced histone H3 acetylation both under static and at 4 hours time point of SS treamtent (Figure 1A). The same result was obtained with an another NOS inhibitor, the 7-Nitroindazole (7N; not shown). This response was associated with an enhanced SS-dependent c-fos expression (Figure 1B). To further investigate the effect of NO on histone acetylation levels, ECs were exposed to a pulse of serum after an overnight starvation, a condition which induces acetylation of histone H3,16 and treated either with the potent NO donor nitroglycerin (NTG) or with the NO second messenger 8Bromide-Cyclic Guanosine Monophospate (8Br-cGMP). Western blot analysis on total histones revealed that NTG and cGMP decreased serum-dependent histone H3 acetylation (Figure 1C) and this was paralleled by the inhibition of serum-induced c-fos expression (Figure 1D). Consistently, treatment of ECs with NTG produced an enhancement in nuclear histone deacetylase activity, as indicated by a HDAC activity assay performed on nuclear extracts (supplemental Figure I).
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NO Induces HDAC4 and HDAC5 Nuclear Shuttling in HUVECs
To clarify NO-dependent mechanism involved in the regulation of gene expression under SS conditions, the subcellular localization of class I and II HDACs was investigated by immunofluorescence. In response to 4 hours of SS class II HDAC4 translocated to the nucleus of ECs, to return in the cytoplasm at 8 hours time point (Figure 2A). Interestingly, this phenomenon was inhibited by SMT treatment (Figure 2B and 2C). The same results were obtained for HDAC5 (not shown), whereas other HDACs tested (1, 2, 3, 6, 7, 8, 9) did not show any change in their cellular localization (not shown). To further verify NO effect on the subcellular localization of class II HDAC4 and 5, ECs were treated either with diethylenetriamine/nitric oxide adduct (DETA/NO), to allow a constant NO release, or with 8BR-cGMP for 1 to 8 hours, and immunofluorescence analysis was performed. As shown in Figure 2D, HDAC4 shuttled to the nucleus at 1 hour of treatment, to return in the cytoplasm at 8 hours time point. HDAC5, which had a more diffuse localization under basal conditions, showed a nuclear enrichment on DETA/NO or 8Bromide-cGMP exposure (supplemental Figure II). Interestingly, in cells treated with 8Br-cGMP, HDAC4 and 5 begin to exit from the nucleus at 4 hours time point. cGMP is metabolized by phosphodiesterases17 (PDEs), and PDE5 has a major role in the pathophysiology of the cardiovascular system.18,19 Treatment of ECs with the PDE5 inhibitor Zaprinast allowed a nuclear enrichment in HDAC4 protein levels at 4 hours of cGMP treatment, as demonstrated by Western blot on ECs nuclear extracts (Figure 3B). In addition, blocking either Erk or p38 kinase, the latter being involved in SS-induced histone acetylation,12 did not interfere with NO-dependent HDAC4 nuclear shuttling; indeed, inhibiting cGMP-activated protein kinase G (PKG) completely abolished HDAC4 nuclear translocation (see supplemental Results and Figure III), addressing a specific role to the NO / cGMP / PKG axis in modulating class II HDACs subcellular localization. Similar results were obtained with HDAC5 (not shown).
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NO Induces Class II HDACs-Dependent Histone Deacetylase Activity in ECs
A series of experiment were performed to investigate whether NO-stimulated histone deacetylation relied on class II HDACs activity. HUVECs were starved for an overnight and the day after were shifted for 1 hour to complete medium supplemented with 10% FBS with or without NTG, in the presence or absence of histone deacetylase inhibitors. As shown in Figure 3C, left panel, NTG significantly decreased serum-dependent histone H3 acetylation. Treatment of ECs with the selective class II HDACs inhibitor MC156820 rescued NTG-dependent histone H3 deacetylation to above control level, whereas the inhibition of class I HDAC1, 2, and 3 activity by the specific inhibitor MS27–27521 had a minor effect. As expected, TSA, which inhibits either class I or class II HDACs, induced a hyperacetylation of histone H3. Altogether, these data suggest an important role for NO in the regulation of histone acetylation by modulating the activity of class II HDACs complexes in ECs.
PP2A Mediates NO-Dependent HDAC4 Nuclear Shuttling
When phosphorylated by calcium-calmodulin dependent kinases (CaMKs) class II HDACs 4 and 5 localize in the cytoplasm bound to the 14-3-3 chaperonins and shuttle to the nucleus in their unphosphorylated form.22 To investigate whether CaMKs may have a role in retaining class II HDACs in the cytoplasm of ECs, HUVECs were exposed to the CaMK inhibitor KN93, in the absence of NO donors. Immunofluorescence analysis showed that KN93 treatment was sufficient to induce HDAC4 nuclear translocation (see supplemental Figure IV), indicating that a CaMkinase activity is required to allow HDAC4 cytosolic retention.
It has been recently demonstrated that CaMKIV and the protein phosphatase PP2A play a role in regulating HDAC5 subcellular localization,23 hence we hypothesized that a PP2A-related activity was involved in NO-dependent HDAC4 nuclear shuttling. To this aim, HUVECs were infected with an adenovirus encoding for the viral small-t antigen oncoprotein, a well known PP2A inhibitor,24 and after 72 hours were treated for 1 to 4 hours with DETA/NO. Phosphatase activity determined on total cell extracts increased in control cells after 1 hour of exposure to DETA/NO, however it was abrogated either in cells expressing small-t antigen or in the presence of the PP2A inhibitor okadaic acid (OA) (Figure 4A and supplemental Figure V). Consistently, NTG also induced an increase in phosphatase activity, which was abolished by OA (Figure 4A and supplemental Figure V). Therefore, to examine whether PP2A was involved in NO-dependent nuclear translocation of HDAC4, HUVECs were stably transfected with the viral oncoprotein small-t.15 Immunofluorescence analysis showed that in mock-transfected cells NTG induced HDAC4 nuclear localization, whereas cells expressing small-t antigen retained HDAC4 in the cytoplasm, suggesting an involvement of PP2A in the regulation of the nuclear shuttling of this enzyme (Figure 4B).
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NO Increases PP2A Binding to HDAC4
To investigate whether NO stimulated the physical association between HDAC4 and PP2A, 293 cells were stably transfected with a doxycyclin inducible HDAC4-Flag fusion protein. 293 cells transfected with the empty vector and induced with doxycyclin were used as control. Western blot analysis on fractionated cellular extracts confirmed the effect of NO on HDAC4 nuclear shuttling also in this cellular model (Figure 5A). Purification of the recombinant protein onto an anti-Flag antibody column and Western blot analysis revealed that in absence of NO, HDAC4-Flag was predominantly associated with the phosphorylated form of CaMKIV25 (p-CaMKIV) and 14-3-3, whereas it was only slightly bound to PP2A, MEF-2,22 HDAC3,26 and HDAC527 (Figure 5B).
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NO treatment caused the dephosphorylation of CaMKIV, coincidently to 14-3-3 detachment, PP2A binding to HDAC4 (see also Figure 6), and the recruitment of MEF-2, HDAC3, and HDAC5 (Figure 5B). PP1 phosphatase was undetectable either in presence or in absence of DETA/NO (not shown). In this condition, HDAC4-associated phosphatase activity was increased on NO exposure, as assessed by an in vitro assay performed by using the recombinant HDAC4-Flag purified from 293 cells untreated or treated with DETA/NO; this activity was inhibited by okadaic acid, either in the presence or absence of NO (Figure 5D).
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Discussion |
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SS exerts a pivotal role in the control of vascular cell structure and function, including the regulation of vascular tone, vessel wall remodeling, and homeostasis, and it is well established that NO is a crucial mediator of some SS properties, including the inhibition of apoptosis and the prevention of atherogenesis.1 NO transcriptional effects have been extensively investigated in different cell types and tissues, including the nervous system,28 the myoblasts,29 and the heart.30,31 In endothelial cells, NO has been described to globally downmodulate gene expression,3 and one of the postulated mechanisms evokes the involvement of DNA methylation, which represses gene expression by favoring a close chromatin conformation.32 The present report describes that the inhibition of NOS prolongs either SS-dependent histone H3 acetylation and c-fos expression (Figure 1A and 1B). The failure of nitric oxide inhibition to produce an increase in H3 acetylation on K14 at earlier time points may be attributable either to the high histone acetyltransferase activity12 or to the low HDAC activity present in endothelial cells at 1 hour of SS exposure. Therefore, the inhibition of histone deacetylase activity is likely to be undetectable at this time point. When histone acetylation begins to decrease, because of both a decrease in HAT activity and an increase in histone deacetylase activity, the inhibition of the latter, by means of nitric oxide inhibitors, becomes visible as an enhacement in histone H3 acetylation. Consistently, it is highly probable that c-fos expression failed to be increased by NOS inhibitors in static conditions because of the requirement of SS-dependent signal transduction pathway to open the chromatin in its promoter region.12 Thus, the genome wide effect of NOS inhibition on histone acetylation may not be paralleled by an effect at gene-specific level, at least in certain conditions. The direct exposure of ECs to the NO donors or to the NO metabolite cGMP abolished the serum induced histone H3 acetylation and c-fos expression (Figure 1C and 1D), indicating a possible direct effect of NO in modulating chromatin structure. In our experimental conditions, ECs nuclear histone deacetylase activity increased after 1 hour of treatment with NTG (supplemental Figure I). These experiments suggested that NO may directly regulate HDACs function. Indeed, we found that SS induced class II HDAC4 (and HDAC5, not shown) nuclear shuttling at 4 hours of treatment and this phenomenon was completely abolished in the presence of the NOS inhibitor SMT (Figure 2A and 2B). Consistently, HDAC4 and 5 shuttled from the cytoplasm to the nucleus of ECs either in the presence of DETA/NO or in the presence of cGMP. This phenomenon was time-dependent, being evident at 1 hour of treatment to decline at 8 hours (Figure 2D). The discrepancy in the timing of class II HDACs nuclear translocation in ECs treated directly with NO donors or exposed to SS (1 hour versus 4 hour) is likely attributable to the requirement of the multiple pathways activated by SS to exert its chromatin remodelling function.12 Moreover, although it is well established that NO production rapidly increase after SS exposure,33 its effects on ECs epigenome may be evident at later time points. Consistent with this hypothesis, it has been described that exposure to shear stress increases NO production in ECs in a biphasic manner.34 An initial rapid NO release occurs at the onset of flow, followed by a less rapid, but sustained production. This constant NO release may allow the NO-dependent class II HDACs nuclear translocation which, in turn, accounts for the regulation of long-term SS-dependent gene expression. In light of this consideration, we reasoned that the transient effect of NO and cGMP relied on the activation of PDEs, which are known to hydrolyze cGMP.17 Indeed, exposure of ECs to Zaprinast, prolonged HDAC4 retention in ECs nuclei, thus supporting our hypothesis of an involvement of PDEs in the nuclear export of class II HDACs (Figure 3B).
SS and NO may activate multiple signals, like p3835 and Erk pathways,36 which are known to be involved in the NO-dependent regulation of cell proliferation and apoptosis. Exposure of ECs to DETA/NO in the presence or absence of Erk and p38 inhibitors had no effect on HDAC4 nuclear translocation, while inhibiting PKG caused HDAC4 cytosolic retention (supplemental Figure III), suggesting an important role for the NO/cGMP/PKG signal transduction pathway in mediating class II HDACs nuclear shuttling in endothelial cells.
A large body of literature assesses that members of the class I and class II HDAC associate with corepressors in a macromolecular complex that mediates deacetylation of histones and repression of transcription.26 Specifically, class II HDACs may function as a structural platform in the context of this chromatin remodeling machinery, where class I histone deacetylases retains the major functional role. In our experiments, however, we found that NO stimulates the histone deacetylase activity of class II HDACs (Figure 3C), addressing a specific role to these enzymes in NO-dependent gene expression and modulation of chromatin structure. Class II HDACs are nuclear in their unphosphorylated form22 and a physical association between HDAC5 and PP2A has also been reported.23 According to these evidences, we demonstrated that HDAC4 (and HDAC5, not shown) failed to localize to the nucleus of ECs in the presence of small-t antigen (Figure 4B), a well known inhibitor of PP2A,24 which also counteracted NO-induced increase of cellular phosphatase activity (Figure 4A). Moreover, we show that PP2A was bound to HDAC4 and HDAC5 in a multiprotein complex, by using 293 cells stably expressing an HDAC4-Flag fusion protein (Figure 5).
The molecular mechanism by which NO may activate PP2A remains to be clarified. The members of PP2A family of phosphatases have a trimeric structure, constituted by a scaffold subunit (the A subunit or PR65) a catalytic subunit (PP2Ac) and a regulatory subunit, which may differ according to the cellular compartment or target.37 Hypothetically, NO may cause a posttranslational modification of PP2A, which accounts for its association with HDAC4. In this regard, it has been demonstrated that phosphorylation of the PP2A regulatory subunit PR61
may either change PP2A substrate affinity38 or enhance the trimer overall activity.39 Our evidence that NO-dependent HDAC4 nuclear translocation involves, at least in part, calcium release (supplemental Figure IV) suggests that a Ca2+-dependent PP2A regulatory subunit40 may associate to and dephosphorylate HDAC4 in response to NO. Mass spectrometric analysis of HDAC4-Flag bound complex will help to identify which regulatory partner enters the PP2A trimer in response to NO.
In conclusion, here we show, for the first time, that nitric oxide induces class II HDAC4 and HDAC5 nuclear shuttling via PP2A activation and provide mechanistic insights into the NO-dependent regulation of gene expression through the regulation of chromatin folding. Although further experiments have to be performed to dissect the molecular mechanisms underlying the NO-induced activation of this chromatin modifier complex, our work may be relevant for a better understanding of the pathogenesis of NO-deficient diseases, like atherosclerosis or inherited pathologies such as duchenne muscular dystrophy.21 According to our results, in fact, a model for NO-dependent class II HDACs nuclear shuttling may be hypothesized (Figure 7). In physiological conditions, NO activates a specific PP2A-related activity which dephosphorylates both CamkIV and class II HDACs, allowing HDACs nuclear translocation and chromatin remodeling. When NO production is impaired, this pathway becomes uneffective, class II HDACs remain in the cytosol contributing to the hyperacetylation of histones or other HDACs target proteins, leading to the disregulation of NO-dependent gene expression.
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Acknowledgments |
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Sources of Funding
This work was supported in part by grant FIRB # RBLA035A4X-1-FIRB to M.C.C., AIRC regional grant to C.G., UE FP6 grant # UE-LHSB-CT-04-502988 to M.C.C., AFM grant # 12042 to C.G.
Disclosures
None.
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Footnotes |
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*The first two authors contributed equally to this study.
Original received December 22, 2006; resubmission received June 5, 2007; revised resubmission received October 12, 2007; accepted October 22, 2007.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
1. Boo YC, Jo H. Flow-dependent regulation of endothelial nitric oxide synthase: role of protein kinases. Am J Physiol Cell Physiol. 2003; 285: C499–C508.
2. Braam B, de Roos R, Bluysen H, Kemmeren P, Holstege F, Joles JA, Koomans H. Nitric oxide-dependent and nitric oxide-independent transcriptional responses to high shear stress in endothelial cells. Hypertension. 2005; 45: 672–680.
3. Braam B, De Roos R, Dijk A, Boer P, Post JA, Kemmeren PP, Holstege FC, Bluyssen HA, Koomans HA. Nitric oxide donors induces temporal and dose-dependent reduction of gene expression in human endothelial cells. Am J Physiol Heart Circ Physiol. 2004; 287: H1877–H1986.
4. Allison CD, Jenuwein T. Translating the histone code Science. 2001; 293: 1074–1080.
5. Sengupta N, Seto E. Regulation of histone deacetylase activity. J Cell Biochem. 2004; 93: 57–67.[CrossRef][Medline] [Order article via Infotrieve]
6. Wade PA. Transcriptional control at regulatory checkpoints by histone deacetylase: molecular connections between cancer and chromatin. Human Molecular Genetics. 2001; 10: 693–698.
7. Laird PW. Cancer epigenetics. Human Molecular Genetics. 2005; 14: R65–R76.
8. Yang X-J, Gregoire S. Class II Histone Deacetylases: frome sequence to function, regulation and clinical implication. Mol Cell Biol. 2005; 25: 2873–2884.
9. Zeng L, Xiao Q, Margariti A, Zhang Z, Zampetaki A, Patel S, Capogrossi MC, Hu Y, Xu Q. HDAC3 is crucial in shear- and VEGF- induced stem cell differentiation towards endothelial cells. J Cell Biol. 2006; 174: 1059–1069.
10. Rossig L, Urbich C, Bruhl T, Dernbach E, Heeschen C, Chavakis E, Sasaki K, Aicher D, Diehl F, Seeger F, Potente M, Aicher A, Zanetta L, Dejana E, Zeiher AM, Dimmeler S. Histone deacetylase activity is essential for the expression of HoxA9 and for andothelial commitment of endothelial progenitor cells. J Exp Med. 2005; 201: 1825–1835.
11. Chang S, Young BD, Li S, Qi X, Richardson JA, Olson EN. Histone deacetylase 7 maintains vascular integrity by repressing matrix metalloproteinase 10. Cell. 2006; 126: 321–334.[CrossRef][Medline] [Order article via Infotrieve]
12. Illi B, Nanni S, Scopece A, Farsetti A, Biglioli P, Capogrossi MC, Gaetano C. Shear stress-mediated chromatin remodelling provides molecular basis for blood flow-dependent regulation of gene expression Circ Res. 2003; 93: 155–161.
13. Illi B, Scopece A, Nanni S, Farsetti A, Morgante L, Biglioli P, Capogrossi MC, Gaetano C. Epigenetic histone modification and cardiovascular lineage programming in mouse embryonic stem cells exposed to laminar shear stress. Circ Res. 2005; 96: 501–508.
14. Dimmeler S, Fleming I, Fisslthaler B, Hermann C, Busse R, Zeiher AM. Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature. 1999; 399: 601–605.[CrossRef][Medline] [Order article via Infotrieve]
15. Cicchillitti L, Fasanaro P, Biglioli P, Capogrossi MC, Martelli F. Oxidative stress induces protein phosphatase 2A-dependent dephosphorylation of the pocket proteins pRb, p107, and p130. J Biol Chem. 2003; 278: 19509–19517.
16. Sassone Corsi P, Mizzen CA, Cheung P, Crosio C, Monaco L, Jacquot S, Hanauer A, Allis CD. Requirement of Rsk-2 for epidermal growth factor-activated phosphorylation of histone H3. Science. 1999; 285: 886–891.
17. Omori K, Kotera J. Overview of PDEs and their regulation. Circ Res. 2007; 100: 309–327.
18. Stehlik J, Movsesia MA. Inhibitors of cyclic nucleotide phosphodiesterase 3 and 5 as therapeutic agents in heart failure. Expert Opin Investig Drugs. 2006; 15: 733–742.[CrossRef][Medline] [Order article via Infotrieve]
19. Munzel T, Daiber A, Mulsch A. Explaining the phenomenon of nitrate tolerance. Circ Res. 2005; 97: 618–628.
20. Mai A, Massa S, Pezzi R, Simeoni S, Rotili D, Nebbioso A, Scognamiglio A, Altucci L, Loidl P, Brosch G. Class II (IIa)-selective histone deacetylase inhibitors. 1. Synthesis and biological evaluation of novel (aryloxopropenyl)pyrrolyl hydroxyamides. J Med Chem. 2005; 48: 3344–3353.[CrossRef][Medline] [Order article via Infotrieve]
21. Minetti GC, Colussi C, Adami R, Serra C, Mozzetta C, Parente V, Fortuni S, Straino S, Sampaolesi M, Di Padova M, Illi B, Gallinari P, Steinkuhler C, Capogrossi MC, Sartorelli V, Bottinelli R, Gaetano C, Puri PL. Functional and morphological recovery of dystrophic muscles in mice treated with deacetylase inhibitors. Nat Med. 2006; 12: 1147–1150.[CrossRef][Medline] [Order article via Infotrieve]
22. McKinsey TA, Zhang CL, Olson EN. Identification of a signal-responsive nuclear export sequence in class II histone deacetylases. Mol Cell Biol. 2001; 21: 6312–6321.
23. Sucharov CC, Langer S, Bristow M, Leinwand L. Shuttling of HDAC5 in H9C2 cells regulates YY1 function through CaMKIV/PKD and PP2A. Am J Physiol Cell Physiol. 2006; 291: C1029–C1037.
24. Arroyo JD, Hahn WC. Involvement of PP2A in viral and cellular transformation. Oncogene. 2005; 24: 7746–7755.[CrossRef][Medline] [Order article via Infotrieve]
25. Miska EA, Langley E, Wolf D, Karlsson C, Pines J, Kouzarides T. Differential localization of HDAC4 orchestrates muscle differentiation. Nucleic Acid Res. 2001; 276: 3439–3447.
26. Fischle W, Dequiedt F, Hendzel MJ, Guenther MG, Lazar MA, Voelter W, Verdin E. Enzymatic activity associated with class II HDACs is dependent on a multiprotein complex containing HDAC3 and SMRT/N-CoR. Mol Cell. 2002; 9: 45–57.[CrossRef][Medline] [Order article via Infotrieve]
27. Kang JS, Alliston T, Delston R, Derynck R. Repression of Runx2 function by TGF-beta through recruitment of class II histone deacetylases by Smad3. EMBO J. 2005; 24: 2543–2555.[CrossRef][Medline] [Order article via Infotrieve]
28. Riccio A, Alvania RS, Lonze BE, Ramanan N, Kim T, Huang Y, Dawson TM, Snyder SH, Ginty DD. A nitric oxide signaling pathway controls CREB-mediated gene expression in neurons. Mol Cell. 2006; 21: 283–294.[CrossRef][Medline] [Order article via Infotrieve]
29. Pisconti A, Brunelli S, Di Padova M, De Palma C, Deponti D, Baesso S, Sartorelli V, Cossu G, Clementi E. Follistatin induction by nitric oxide through cyclic GMP: a tightly regulated signaling pathway that controls myoblast fusion. J Cell Biol. 2006; 172: 233–244.
30. Suzuki YJ, Nagase H, Day RM, Das DK. GATA-4 regulation of myocardial survival in the preconditioned heart. J Mol Cell Cardiol. 2004; 37: 1195–1203.[CrossRef][Medline] [Order article via Infotrieve]
31. Kuwabara M, Kakinuma Y, Ando M, Katare RG, Yamasaki S, Doi Y, Sato T. Nitric oxide stimulates vascular endothelial growth factor production in cardiomyocytes involved in angiogenesis. J Physiol Sci. 2006; 56: 95–101.[CrossRef][Medline] [Order article via Infotrieve]
32. Hmadcha A, Bedoya FJ, Sobrino F, Pintado E. Methylation-dependent gene silencing induced by interleukin 1beta via nitric oxide production. J Exp Med. 1999; 190: 1595–1604.
33. Ohno M, Gibbons GH, Dzau VJ, Cooke JP. Shear stress elevates endothelial cGMP. Role of potassium channel and G protein coupling. Circulation. 1993; 88: 193–197.
34. Kuchan MJ, Frangos JA. Role of calcium and calmodulin in flow-induced nitric oxide production in endothelial cells. Am J Physiol. 1994; 35: C628–C636.
35. Ptasinska A, Wang S, Zhang J, Wesley RA, Danner RL. Nitric oxide activation of peroxisome proliferator-activated receptor gamma through a p38 MAPK signaling pathway. FASEB J. 2007; 21: 950–961.
36. Zuckerbraun BS, Stoyanovsky DA, Sengupta R, Shapiro RA, Ozanich BA, Rao J, Barbato JE, Tzeng E. Nitric oxide-induced inhibition of smooth muscle cell proliferation involves S-nitrosation and inactivation of RhoA. Am J Physiol Cell Physiol. 2007; 292: C824–C831.
37. Janssens V, Goris J. Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatase implicated in cell growth and signalling. Biochem J. 2001; 353: 417–439.[CrossRef][Medline] [Order article via Infotrieve]
38. Usui H, Inoue R, Tanabe O, Nishito Y, Shimizu M, Hayashi H, Kagamiyama H, Takeda M. Activation of protein phosphatase 2A by cAMP-dependent protein kinase-catalyzed phosphorylation of the 74-kDa B'' (delta) regulatory subunit in vitro and identification of the phosphorylation sites. FEBS Lett. 1998; 430: 312–316.[CrossRef][Medline] [Order article via Infotrieve]
39. Xu Z, Williams BRG. The B56alpha regulatory subunit of protein phosphatase 2A is a target for regulation by double-stranded RNA-dependent protein kinase PKR. Mol Cell Biol. 2000; 20: 5285–5299.
40. Janssens V, Jordens J, Stevens I, Van Hoof C, Martens E, De Smedt H, Engelborghs Y, Waelkens E, Goris J. Identification and functional analysis of two Ca2+-binding EF-hand motifs in the B''/PR72 subunit of protein phosphatase 2A. J Biol Chem. 2003; 278: 10697–10706.
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