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Up-regulation of human inducible nitric oxide synthase by p300 transcriptional complex

Guo, Z and Zheng, L and Liao, X and Geller, D (2016) Up-regulation of human inducible nitric oxide synthase by p300 transcriptional complex. PLoS ONE, 11 (1).

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p300, a ubiquitous transcription coactivator, plays an important role in gene activation. Our previous work demonstrated that human inducible nitric oxide synthase (hiNOS) expression can be highly induced with the cytokine mixture (CM) of TNF-α + IL-1β + IFN-γ. In this study, we investigated the functional role of p300 in the regulation of hiNOS gene expression. Our initial data showed that overexpression of p300 significantly increased the basal and cytokine-induced hiNOS promoter activities in A549 cells. Interestingly, p300 activated cytokine-induced hiNOS transcriptional activity was completely abrogated by deleting the upstream hiNOS enhancer at -5 kb to -6 kb in the promoter. Furthermore, p300 over-expression increased cytokine-induced transcriptional activity on a heterologous minimal TK promoter with the same hiNOS enhancer. Site-directed mutagenesis of the hiNOS AP-1 motifs revealed that an intact upstream (-5.3kb) AP-1 binding site was critical for p300 mediated cytokine-induced hiNOS transcription. Furthermore, our ChIP analysis demonstrated that p300 was binding to Jun D and Fra-2 proteins at -5.3 kb AP-1 binding site in vivo. Lastly, our 3C assay was able to detect a long DNA loop between the hiNOS enhancer and core promoter site, and ChIP loop assay confirmed that p300 binds to AP-1 and RNA pol II proteins. Overall, our results suggest that coactivator p300 mediates cytokine-induced hiNOS transactivation by forming a distant DNA loop between its enhancer and core promoter region.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Guo, Zzhg1@pitt.eduZHG1
Zheng, L
Liao, Xxil57@pitt.eduXIL57
Geller, Ddageller@pitt.eduDAGELLER
ContributionContributors NameEmailPitt UsernameORCID
Date: 11 January 2016
Date Type: Publication
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Journal or Publication Title: PLoS ONE
Volume: 11
Number: 1
DOI or Unique Handle: 10.1371/journal.pone.0146640
Institution: University of Pittsburgh
Schools and Programs: School of Medicine > Medicine
School of Medicine > Surgery
Refereed: Yes
Date Deposited: 23 Aug 2016 14:56
Last Modified: 30 Mar 2021 11:55


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