Cloning and characterization of the murine β 3 integrin gene promoter: Identification of an interleukin-4 responsive element and regulation by STAT-6

Kevin P. McHugh, Sohei Kitazawa, Steven L. Teitelbaum, F. Patrick Ross

Research output: Contribution to journalArticle

30 Scopus citations

Abstract

Expression of the α vβ 3 integrin by murine bone marrow macrophages is regulated by cytokines such as IL-4 and GM-CSF through transcriptional activation of the β 3 subunit gene. To characterize the molecular mechanisms by which such regulation occurs, we isolated the murine β 3 integrin promoter. To this end, we first cloned a full length β 3 cDNA and used the 5′UTR and leader peptide coding sequence to identify genomic clones containing the β 3 promoter region. The transcriptional start site, identified by primer extension and S1 nuclease assay, is 34 nt upstream of the translation initiation codon. A 1.1 kb fragment of the promoter region drives IL-4 responsive transcription in transiently transfected murine bone marrow macrophages. Deletion analysis of the β 3 promoter indicates the IL-4 responsive element lies between -465 to -678 nt relative to the transcriptional start site. This promoter fragment contains two overlapping STAT consensus recognition sites and nuclear extracts from BMMs contain an IL-4-inducible DNA binding factor, identified by super shift analysis, as STAT-6. Furthermore, an oligonucleotide which includes the two STAT recognition sites residing in the IL-4 responsive region of the β 3 promoter, competes for STAT-6 binding. Confirming IL-4 induction of the integrin subunit is specifically mediated by STAT-6, β 3 mRNA is not enhanced in BMMs derived from STAT-6 deleted mice, which however, retain their capacity to respond to GM-CSF.

Original languageEnglish
Pages (from-to)320-332
Number of pages13
JournalJournal of cellular biochemistry
Volume81
Issue number2
DOIs
StatePublished - Apr 25 2001

Keywords

  • Bone marrow macrophages
  • Interleukin 4
  • Promoter analysis
  • STAT-6
  • β integrin

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