Role of the membrane anchoring and cytoplasmic domains in intracellular transport and localization of viral glycoproteins

1999 ◽  
Vol 77 (3) ◽  
pp. 165-178 ◽  
Author(s):  
Kakoli Ghosh ◽  
Hara P Ghosh
Keyword(s):  
Nitric Oxide ◽  
Intracellular Transport ◽  
Transendothelial Migration ◽  
Interleukin 8 ◽  
Nitric Oxide Donor ◽  
Luciferase Reporter ◽  
Nuclear Factor ◽  
Reporter Construct ◽  
Luciferase Reporter Construct ◽  
Exogenous Nitric Oxide

Nuclear factor-κB (NF-κB) binds to nucleotide sequences between -80 and -70 bp upstream of the transcriptional start site in the interleukin-8 (IL-8) promoter and is crucial for transcription of the IL-8 gene. We showed that exogenous nitric oxide in the form of a nitric oxide donor significantly reduced IL-8 mRNA in cytokine-activated ECV304. Similarly, nitric oxide significantly reduced migration of polymorphonuclear neutrophils through cytokine-activated ECV304 monolayers, an IL-8-dependent process. Using a luciferase reporter construct containing the NF-κB site of the IL-8 gene, we showed that exposing cytokine-activated ECV304 to exogenous nitric oxide resulted in significant reduction of NF-κB binding. Follow-up studies using a luciferase reporter construct possessing a mutated NF-κB site confirmed that the luciferase activity observed in the NF-κB reporter resulted from NF-κB binding. These studies demonstrate that nitric oxide, supplied exogenously into reactions containing activated endothelium, down-regulates pro-inflammatory activity, such as the secretion of chemokines, and functional activity, such as transendothelial migration of neutrophils. Key words: interleukin-8, nuclear factor κ B, transendothelial migration, nitric oxide.

Nitric oxide regulates interleukin-8 gene expression in activated endothelium by inhibiting NF-κB binding to DNA: effects on endothelial function

1999 ◽  
Vol 77 (3) ◽  
pp. 201-208 ◽  
Author(s):  
Alph A Fowler, III ◽  
Bernard J Fisher ◽  
Lori B Sweeney ◽  
Timothy J Wallace ◽  
Ramesh Natarajan ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Transendothelial Migration ◽  
Interleukin 8 ◽  
Polymorphonuclear Neutrophils ◽  
Nitric Oxide Donor ◽  
Luciferase Reporter ◽  
Nuclear Factor ◽  
Reporter Construct ◽  
Luciferase Reporter Construct ◽  
Exogenous Nitric Oxide

Nuclear factor-kappaB (NF-κB) binds to nucleotide sequences between -80 and -70 bp upstream of the transcriptional start site in the interleukin-8 (IL-8) promoter and is crucial for transcription of the IL-8 gene. We showed that exogenous nitric oxide in the form of a nitric oxide donor significantly reduced IL-8 mRNA in cytokine-activated ECV304. Similarly, nitric oxide significantly reduced migration of polymorphonuclear neutrophils through cytokine-activated ECV304 monolayers, an IL-8-dependent process. Using a luciferase reporter construct containing the NF-κB site of the IL-8 gene, we showed that exposing cytokine-activated ECV304 to exogenous nitric oxide resulted in significant reduction of NF-κB binding. Follow-up studies using a luciferase reporter construct possessing a mutated NF-κB site confirmed that the luciferase activity observed in the NF-kappaB reporter resulted from NF-κB binding. These studies demonstrate that nitric oxide, supplied exogenously into reactions containing activated endothelium, down-regulates pro-inflammatory activity, such as the secretion of chemokines, and functional activity, such as transendothelial migration of neutrophils.Key words: interleukin-8, nuclear factor κ B, transendothelial migration, nitric oxide.

scholarly journals In Vivo Imaging of Transiently Transgenized Mice with a Bovine Interleukin 8 (CXCL8) Promoter/Luciferase Reporter Construct

PLoS ONE ◽  
2012 ◽  
Vol 7 (6) ◽  
pp. e39716 ◽  
Author(s):  
Fabio Franco Stellari ◽  
Valentina Franceschi ◽  
Antonio Capocefalo ◽  
Marcello Ronchei ◽  
Fabrizio Facchinetti ◽  
...  
Keyword(s):  
In Vivo Imaging ◽  
Interleukin 8 ◽  
Luciferase Reporter ◽  
Reporter Construct ◽  
Luciferase Reporter Construct

Myocardial relaxant effect of exogenous nitric oxide in isolated ejecting hearts

1994 ◽  
Vol 266 (5) ◽  
pp. H1699-H1705 ◽  
Author(s):  
R. Grocott-Mason ◽  
S. Fort ◽  
M. J. Lewis ◽  
A. M. Shah
Keyword(s):  
Nitric Oxide ◽  
Sodium Nitroprusside ◽  
Coronary Flow ◽  
Systolic Function ◽  
Myocardial Contraction ◽  
Left Ventricular ◽  
Nitric Oxide Donor ◽  
Vasodilator Activity ◽  
Exogenous Nitric Oxide ◽  
Whole Heart

In isolated myocytes and papillary muscles, both nitric oxide, acting through guanosine 3',5'-cyclic monophosphate (cGMP), and cGMP analogues exert a novel effect on myocardial contraction, influencing mainly the onset of relaxation. We studied the effect of the exogenous nitric oxide donor, sodium nitroprusside (0.1-10 microM), in isolated ejecting guinea pig hearts at constant filling pressure, afterload, and heart rate to identify its direct myocardial effects in the whole heart. Sodium nitroprusside induced concentration-dependent increases in coronary flow as well as premature and faster early left ventricular (LV) pressure decline, but did not change end-diastolic or peak LV pressure or peak rate of rise of LV pressure. There was no correlation between changes in coronary flow and LV pressure decline. Sodium nitroprusside effects were inhibited by hemoglobin, which inactivates nitric oxide. The cGMP-independent vasodilator nicardipine also increased coronary flow but did not influence early LV pressure fall. Thus exogenous nitric oxide exerts novel direct myocardial relaxant effects in the isolated ejecting heart, independent of its known vasodilator activity, and without compromising systolic function.

Mapping and functional analysis of an instability element in phosphoenolpyruvate carboxykinase mRNA

2000 ◽  
Vol 279 (5) ◽  
pp. F866-F873 ◽  
Author(s):  
Omar F. Laterza ◽  
Lynn Taylor ◽  
Shashikala Unnithan ◽  
Ly Nguyen ◽  
Norman P. Curthoys
Keyword(s):  
Phosphoenolpyruvate Carboxykinase ◽  
Luciferase Activity ◽  
Specific Protein ◽  
Luciferase Reporter ◽  
Reporter Construct ◽  
Luciferase Reporter Construct ◽  
Functional Studies ◽  
Renal Gluconeogenesis ◽  
Shift Analysis ◽  
Nontranslated Region

Phosph oenolpyruvate carboxykinase (PEPCK) is a key regulatory enzyme of renal gluconeogenesis. The 3′-nontranslated region of the PEPCK mRNA contains an instability element that facilitates its rapid turnover and contributes to the regulation of PEPCK gene expression. Such processes are mediated by specific protein-binding elements. Thus RNA gel shift analysis was used to identify proteins in rat renal cortical cytosolic extracts that bind to the 3′-nontranslated region of the PEPCK mRNA. Deletion constructs were then used to map the binding interactions to two adjacent RNA segments (PEPCK-6 and PEPCK-7). However, competition experiments established that only the binding to PEPCK-7 was specific. Functional studies were performed by cloning similar segments in a luciferase reporter construct, pLuc/Zeo. This analysis indicated that both PEPCK-6 and PEPCK-7 segments were necessary to produce a decrease in luciferase activity equivalent to that observed with the full-length 3′-nontranslated region. Thus the PEPCK-7 segment binds a specific protein that may recruit one or more proteins to form a complex that mediates the rapid decay of the PEPCK mRNA.

Nuclear factor-κB activation by the photochemotherapeutic agent verteporfin

Blood ◽  
2000 ◽  
Vol 95 (1) ◽  
pp. 256-262 ◽  
Author(s):  
D. J. Granville ◽  
C. M. Carthy ◽  
H. Jiang ◽  
J. G. Levy ◽  
B. M. McManus ◽  
...  
Keyword(s):  
Stress Responses ◽  
Nuclear Factor Κb ◽  
Light Irradiation ◽  
Luciferase Reporter ◽  
Nuclear Factor ◽  
Luciferase Reporter Construct ◽  
Nuclear Extracts ◽  
The Status ◽  
General Caspase Inhibitor ◽  
Induced Apoptosis

The nuclear factor-kappa B (NF-κB) gene transactivator serves in the formation of immune, inflammatory, and stress responses. In quiescent cells, NF-κB principally resides within the cytoplasm in association with inhibitory κ (IκB) proteins. The status of IκB and NF-κB proteins was evaluated for promyelocytic leukemia HL-60 cells treated at different intensities of photodynamic therapy (PDT). The action of the potent photosensitizer, benzoporphyrin derivative monoacid ring A (verteporfin), and visible light irradiation were assessed. At a verteporfin concentration that produced the death of a high proportion of cells after light irradiation, evidence of caspase-3 and caspase-9 processing and of poly(ADP-ribose) polymerase cleavage was present within whole cell lysates. The general caspase inhibitor Z-Val-Ala-Asp-fluoromethylketone (ZVAD.fmk) effectively blocked these apoptosis-related changes. Recent studies indicate that IκB proteins may be caspase substrates during apoptosis. However, the level of IκBβ was unchanged for HL-60 cells undergoing PDT-induced apoptosis. IκB levels decreased during PDT-induced apoptosis, though ZVAD.fmk did not affect this change. At a less intensive level of photosensitization, cellular IκB levels were transiently depressed after PDT. At these times, p50 and RelA NF-κB species were increased within nuclear extracts, as revealed by electrophoretic mobility supershift assays. HL-60 cells transiently transfected with a κB-luciferase reporter construct exhibited elevated luciferase activity after PDT or treatment with tumor necrosis factor-, a well-characterized NF-κB activator. Productive NF-κB activation and associated gene transcription may influence the phenotype and behavior of cells exposed to less intensive PDT regimens. However, IκB is not subject to caspase-mediated degradation as a component of PDT-induced apoptosis. (Blood. 2000;95:256-262)

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