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PMID: 8289831 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A unique palindromic element mediates gamma interferon induction of mig gene expression.

Molecular and cellular biology ·Vol. 14 ·No. 2 ·1994-02-00 ·Pages 914-22

Wong P, Severns CW, Guyer NB, Wright TM

Abstract

To define the molecular mechanisms involved in the action of gamma interferon (IFN-gamma), we have analyzed the transcriptional regulation of the mig (monokine induced by gamma interferon) gene, a member of the platelet factor 4-interleukin-8 cytokine family that is expressed in murine macrophages specifically in response to IFN-gamma. Analysis of mig/CAT chimeric constructs transiently transfected into the RAW 264.7 mouse monocytic cell line revealed a unique IFN-gamma-responsive element (gamma RE-1). The sequence of this cis regulatory element defined by deletion analysis contains an imperfect inverted repeat extending 27 bp. Examination of mig/CAT constructs with mutations in gamma RE-1 revealed that the palindromic positions in the element were essential for activity. Consistent with its function as an enhancer, a single copy of gamma RE-1 conferred IFN-gamma inducibility to a heterologous (herpes simplex virus thymidine kinase) promoter. Exonuclease III protection assays demonstrated symmetrical protection of a mig promoter fragment centered about the gamma RE-1 palindromic sequence. Using the gel electrophoretic mobility shift assay, we identified a factor (gamma RF-1) present in nuclear extracts prepared from IFN-gamma-stimulated RAW 264.7 cells which binds to gamma RE-1. The activation of gamma RF-1 occurred rapidly (within 1 min) in response to IFN-gamma and was independent of protein synthesis. Similar to the expression of mig mRNA, the formation of gamma RF-1 was selectively induced by IFN-gamma and not IFN-alpha. The regulation of gene expression through gamma RF-1 and gamma RE-1 may explain the preferential activation of a subset of interferon-inducible genes by IFN-gamma.

Related Genes
mig
MeSH Terms
Animals Base Sequence Cell Line Cell Nucleus/metabolism Chloramphenicol O-Acetyltransferase/biosynthesis,metabolism Cytokines/biosynthesis Cytosol/metabolism DNA Primers Enhancer Elements, Genetic Exodeoxyribonucleases Gene Expression/drug effects Interferon-gamma/pharmacology Mice Molecular Sequence Data Monocytes Polymerase Chain Reaction Promoter Regions, Genetic RNA, Messenger/biosynthesis Recombinant Proteins Repetitive Sequences, Nucleic Acid Simplexvirus/genetics Thymidine Kinase/genetics Transcription, Genetic/drug effects Transfection
Chemicals
Cytokines DNA Primers RNA, Messenger Recombinant Proteins Interferon-gamma Chloramphenicol O-Acetyltransferase Thymidine Kinase Exodeoxyribonucleases exodeoxyribonuclease III
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wong P
Department of Medicine, University of Pittsburgh School of Medicine, Pennsylvania 15261.
Severns C W
Guyer N B
Wright T M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.2.914-922.1994
Published
1994-02-00
Pages
914-22
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358446
Subset
IM
Grants
NCI NIH HHS · CA55333 · United States
External Links
PubMed source DOI: 10.1128/mcb.14.2.914-922.1994 PMC Full Text View journal details
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