Identification of hamster inducible nitric oxide synthase (iNOS) promoter sequences that influence basal and inducible iNOS expression

Omar A. Saldarriaga, Bruno Travi, Goutam Ghosh Choudhury, Peter Melby

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

IFN-γ/LPS-activated hamster (Mesocricetus auratus) macrophages express significantly less iNOS (NOS2) than activated mouse macrophages, which contributes to the hamster's susceptibility to intracellular pathogens. We determined a mechanism responsible for differences in iNOS promoter activity in hamsters and mice. The HtPP (1.2 kb) showed low basal and inducible promoter activity when compared with the mouse, and sequences within a 100-bp region (-233 to -133) of the mouse and hamster promoters influenced this activity. Moreover, within this 100 bp, we identified a smaller region (44 bp) in the mouse promoter, which recovered basal promoter activity when swapped into the hamster promoter. The mouse homolog (100-bp region) contained a cis-element for NF-IL-6 (-153/-142), which was absent in the hamster counterpart. EMSA and su-pershift assays revealed that the hamster sequence did not support the binding of NF-IL-6. Introduction of a functional NF-IL-6 binding sequence into the hamster promoter or its alteration in the mouse promoter revealed the critical importance of this transcription factor for full iNOS promoter activity. Furthermore, the binding of NF-IL-6 to the iNOS promoter (-153/-142) in vivo was increased in mouse cells but was reduced in hamster cells after IFN-γ/LPS stimulation. Differences in the activity of the iNOS promoters were evident in mouse and hamster cells, so they were not merely a result of species-specific differences in transcription factors. Thus, we have identified unique DNA sequences and a critical transcription factor, NF-IL-6, which contribute to the overall basal and inducible expression of hamster iNOS.

Original languageEnglish (US)
Pages (from-to)205-218
Number of pages14
JournalJournal of Leukocyte Biology
Volume92
Issue number1
DOIs
StatePublished - Jul 2012
Externally publishedYes

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Nitric Oxide Synthase Type II
Cricetinae
Interleukin-6
Transcription Factors
Macrophages
Mesocricetus

Keywords

  • C/EBP-β
  • NF-IL-6
  • NOS2
  • Transcriptional regulation

ASJC Scopus subject areas

  • Cell Biology
  • Immunology

Cite this

Identification of hamster inducible nitric oxide synthase (iNOS) promoter sequences that influence basal and inducible iNOS expression. / Saldarriaga, Omar A.; Travi, Bruno; Choudhury, Goutam Ghosh; Melby, Peter.

In: Journal of Leukocyte Biology, Vol. 92, No. 1, 07.2012, p. 205-218.

Research output: Contribution to journalArticle

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