Modulation of AP‐1 activity by nitric oxide (NO) in vitro: NO‐mediated modulation of AP‐1

Abstract

To understand the role of nitric oxide (NO) in controlling the specific DNA-binding activities of transcriptional factors, we investigated the in vitro effect of the NO-donor sodium nitroprusside (SNP) on the AP-1 activity of cultured mouse cerebellar granule cells. A gel-mobility assay showed that SNP inhibited AP-1 activity in the presence, but not the absence of dithiothreitol (DTT). This DTT-dependent inhibition of AP-1 activity by SNP corresponded with the activation of the chemical reactivity of SNP with DTT, which can be monitored by the production of nitrite (NO;). In contrast, diamide, a typical sulfhydryl oxidizing agent, inhibited AP-I activity in the absence of DTT and its inhibitory effect was reversed competitively by DTT. Studies using structurally or functionally related analogues of SNP demonstrated that S-nitrosylation of the AP-1 moiety mediated by some NO-carriers but not by free NO, which can be produced by the chemical reaction of SNP with DTT, was responsible for the inhibition of AP-1 activity, suggesting NO-mediated regulation of the AP-1 transcriptional factor.

Regulation and measurement of oxidative stress in apoptosis

“…These include tissue transglutaminase (Ttg), (Melino et al, 1997), the two cysteine-transcription factors NF kappa B (DelaTorre and DelaTorre), and AP-1 (Tabuchi et al, 1994), both implicated in the regulation of apoptosis, several ion channels (Broillet; Eu and Hart) and the protease calpain (Michetti et al, 1995) (Fig. 3).…”

Regulation of the apoptosis–necrosis switch

“…It affects the cellular gene transcription machinery by inhibiting the DNA-binding activity of nuclear factor kB (NF-kB) (Matthews et al, 1996;Marshall and Stamler, 2001) and activator protein-1 (AP-1) (Tabuchi et al, 1994) or by activation of OxyR (Hausladen et al, 1996). NO nitrosylates thiol groups of cytosolic enzymes such as transglutaminase (Melino et al, 1997;Bernassola et al, 1999;Rossi et al, 2000), AP-1 Melino et al, 2000a), caspases (Dimmeler et al, 1997;Melino et al, 1997), glyceraldehyde-3-phosphate dehydrogenase (Mohr et al, 1996) hemoglobin (Jia et al, 1996;Gow and Stamler, 1998) and skeletal muscle (Stamler and Meissner, 2001;Eu et al, 2003), and it can modulate the biological activity of extracellular molecules such as coagulation factor XIII (Catani et al, 1998).…”

A Sp1 Binding Site of the Tumor Necrosis Factor α Promoter Functions as a Nitric Oxide Response Element

“…Work in this area has focused on relatively large promoter regions that appear to be involved in these NO mediated responses (6 -7, 27). Other studies have identified transcription factors including NF-B (3, 28), AP1 (29), heat shock factor 1 (30), and Oct-1 (31) that undergo increased or decreased DNA binding upon exposure of cells or cell extract to exogenous NO. Taken together, these results suggested that NO signaling cascades extend to the level of gene transcription through multiple promoter regulatory sites.…”