Abstract
A newly described member of the platelet factor 4 family of cytokine genes, mig, is selectively induced by interferon gamma (IFN-gamma), and not IFN-alpha, in the mouse macrophage-like cell line RAW 264.7. Treatment of RAW 264.7 cells with IFN-gamma activated mig gene transcription as determined by nuclear run-on assays. mig genomic clones were isolated, and constructs containing genomic fragments that included the mig promoter region and the CAT reporter gene were prepared. In RAW 264.7 cells transfected with these constructs, CAT activity was found to be selectively induced by IFN-gamma. A 278-bp genomic fragment containing 235 nucleotides 5' of the transcription start site was sufficient for IFN-gamma-selective induction of CAT activity. Analysis of 5' deletion mutants localized a region essential for activation by IFN-gamma to within 64 nucleotides extending from -235 to -172. A genomic fragment containing this sequence was capable of conferring IFN-gamma inducibility to constructs with a heterologous promoter.
Highlights
The mig cDNA clone was obtained by differential screening of a cDNA library prepared from the mouse macrophage-like cell line RAW 264.7 treated with conditioned medium from Con A-stimulated mouse splenocytes
The mig cDNA hybridizes to a 1.6-kb mRNA selectively induced by interferon y (IFN-y) and encodes a new member of the platelet factor 4 family of cytokines [14]
The expression of mig mRNA was potently induced by IFN-y, but not by IFN-a, over a wide concentration range
Summary
The mig (monokine induced by IFN-y) cDNA clone was obtained by differential screening of a cDNA library prepared from the mouse macrophage-like cell line RAW 264.7 treated with conditioned medium from Con A-stimulated mouse splenocytes . Constructs were prepared containing the mig promoter region and 5' flanking sequences inserted into the promoterless CAT reporter plasmid pUMSVOCAT The initial mig/ CAT plasmid contained an 1,160-bp mig genomic fragment extending from nucleotides -1117 to +43 relative to the transcription start site.
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