IFN-γ upregulates expression of the mouse complement C1rA gene in keratinocytes via IFN-regulatory factor-1

Sung June Byun, Ik Soo Jeon, Hyangkyu Lee, Tae Yoon Kim

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We examined the expression of the mouse complement component C1rA (mC1rA) in IFN-γ-stimulated mouse keratinocytes (Pam 212) and found that it was upregulated. To analyze the mechanism involved, we cloned the 2,150 bp 5′-flanking region of mC1rA by the vectorette-PCR technique, and identified the transcription start site of mC1rA by rapid amplification of complementary DNA ends. Analysis of the 5′ sequence revealed putative binding sites for activator protein 1, CCAAT/enhancer binding protein (C/EBP), signal transducer and activator of transcription 1 (STAT-1), IFN-regulatory factor-1 (IRF-1), and others. We detected transcriptional activation dependent on this upstream region in reporter gene assays and Northern blots. To identify the cis-acting regulatory elements involved, we analyzed serial deletion constructs of the promoter using luciferase reporters. The -80 to -19 bp region, which contains a putative IRF-1 binding site, was required for both basal promoter activity and responses to IFN-γ. The use of site-directed point mutations, electrophoresis mobility shift assays, and supershift assays indicated that the putative IRF-1 binding site was essential for both IFN-γ-dependent and -independent transcriptional activity of the mC1rA promoter. We conclude that IFN-γ stimulates mC1rA gene expression via IRF-1 in mouse keratinocytes.

Original languageEnglish
Pages (from-to)1187-1196
Number of pages10
JournalJournal of Investigative Dermatology
Volume127
Issue number5
DOIs
Publication statusPublished - 2007 May

Bibliographical note

Funding Information:
This work was supported by National Research Laboratory (NRL) Grant (M1-0104-00-0266) from the Korean Ministry of Science and Technology. We thank Dr Stuart Yuspa and Professor N. Fusenig for providing Pam 212 and HaCaT cells, respectively.

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Dermatology
  • Cell Biology

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