title: Protein/DNA arrays identify nitric oxide-regulated cis-element and trans-factor activities some of which govern neuroblastoma cell viability.
abstract: Toxic nitric oxide (NO) levels can regulate gene expression. Using a novel protein/DNA array, we show that toxic NO levels regulate the binding of trans-factors to various cis-elements in neuroblastoma cells, including CRE and those recognized by the transcription factors AP1, AP2, Brn-3a, EGR, E2F1 and SP1. Functionality of some of the cis-elements was confirmed by electro mobility shift and reporter assays. Interestingly, CREB, AP-1, Brn-3a, EGR and E2F1 can control mammalian cell viability. NO induced the anti-apoptotic Bcl-2 protein and its mRNA prior to the onset of death of 30-60% of the cells. Promoter analysis of the bcl-2 gene confirmed the involvement of a CRE in NO-dependent bcl-2 transcription. Neuroblastoma cells over-expressing bcl-2 became much more resistant to NO-induced apoptosis; conversely, Bcl-2 knockdown cells were rendered markedly more sensitive to NO. Together these results suggest that Bcl-2 counteracts NO-induced apoptosis in a fraction of the cell population. Thus, NO stimulates the binding of many trans-factors to their cognate cis-elements, some of which can regulate cell viability through transcriptional activation of target genes. Our results emphasize that a DNA/protein array approach can reveal novel, global transcription factor activities stimulated by cell death-regulating molecules.
authors: Cell Death and Human Diseases Group, Division of Genomics and Genetics, Institute of Molecular & Cell Biology, Proteos, 61 Biopolis Drive, Singapore 138673, Republic of Singapore.
authors: Saravanakumar
authors: S
authors: Dhakshinamoorthy
authors: Shiva Ranjani
authors: SR
authors: Sridharan
authors: Lei
authors: L
authors: Li
authors: Poh Yong
authors: PY
authors: Ng
authors: Linda M
authors: LM
authors: Boxer
authors: Alan G
authors: AG
authors: Porter
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D001665:Binding Sites; D045744:Cell Line, Tumor; D002470:Cell Survival; D000242:Cyclic AMP / Q000378:metabolism; D019254:Genes, bcl-2; D006801:Humans; D009447:Neuroblastoma; D009569:Nitric Oxide / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D040081:Protein Array Analysis; D020218:Response Elements; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation
chemical_list: D014157:Transcription Factors; D009569:Nitric Oxide; D000242:Cyclic AMP
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