A novel gene family induced by acute inflammation in endothelial cells.
The aim of this study was to characterise a novel family of inflammatory genes induced by pro-inflammatory cytokines in primary human endothelial cells. Using a genome-wide array screen two previously uncharacterised genes, NLF1 and NLF2 were identified that were upregulated over 30 fold by treatment with interleukin 1beta for 2 h. They were also found to respond to tumour necrosis factor alpha, suggesting a general role in inflammation. Expression of both genes peaked 2 h after addition of interleukin 1beta, with similar kinetics to the fastest nuclear factor kappaB (NF-kappaB) induced genes. The activation of both genes by interleukin 1beta was abrogated by the proteasomal inhibitor, lactacystin which blocks activation of NF-kappaB by preventing IkappaB degradation. Furthermore, two sequences with homology to NF-kappaB binding sites in the promoter of NLF1 were found to be essential for rapid elevation in expression in response to interleukin 1beta. NLF1 and NLF2 transcripts were found predominantly in endothelial cells, and the encoded proteins were localised to the nuclear compartment suggesting a role in the regulation of transcription. Transfection of recombinant NLF into endothelial cells resulted in upregulation of the Rho kinases, Rnd1 and Gem GTPase. We propose that NLF1 and NLF2 belong to a novel gene family encoding nuclear factors with a role in regulating genes which control cellular architecture. This might increase vascular permeability in acute inflammation.
Warton
Kristina
K
Centre for Immunology, University of New South Wales and St. Vincent's Hospital, Darlinghurst NSW 2010, Sydney, Australia.
Foster
Natasha C
NC
Gold
Wendy A
WA
Stanley
Keith K
KK
D000208:Acute Disease; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D018938:Caco-2 Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D002884:Chromosomes, Human, Pair 15 / Q000235:genetics; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D005091:Exons; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D023061:Gene Order; D005796:Genes / Q000235:genetics; D015894:Genome, Human; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000235:genetics; D007375:Interleukin-1 / Q000494:pharmacology; D015850:Interleukin-6 / Q000235:genetics; D007438:Introns; D008565:Membrane Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009687:Nuclear Proteins; D020411:Oligonucleotide Array Sequence Analysis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics
C494527:C2CD4A protein, human; C494528:C2CD4B protein, human; D007375:Interleukin-1; D015850:Interleukin-6; D008565:Membrane Proteins; D016328:NF-kappa B; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; C063552:transcription factor BTF3; D049452:Green Fluorescent Proteins