Interferon-α regulates nuclear translocation and DNA-binding affinity of ISGF3, a multimeric transcriptional activator

D. S. Kessler, S. A. Veals, X. Y. Fu, D. E. Levy

Research output: Contribution to journalArticle

237 Scopus citations

Abstract

The interaction of interferon-α (IFN-α) with a specific cell-surface receptor elicits physiological changes that rely on rapid transcriptional activation of a group of IFN-α-stimulated genes (ISGs). The IFN-stimulated response element (ISRE), a conserved regulatory element of all ISGs, is the target for transcriptional activation by the positive regulator IFN-stimulated gene factor-3 (ISGF3). We reported previously that post-translational activation of ISGF3 in the cytoplasm of IFN-α-treated cells requires two cytoplasmic activities (ISGF3α and ISGF3γ) to produce an ISRE-binding complex that accumulates in the nucleus. In this study, we show that these activities are actually distinct subunits of the ISGF3 complex, which associate through noncovalent interaction. Sedimentation analysis, protein renaturation, and photoaffinity cross-linking of enriched preparations of cytoplasmic ISGF3α and ISGF3γ and of nuclear ISGF3 demonstrated that ISGF3γ was a 48-kD polypeptide with intrinsic, low-affinity DNA-binding activity. Four polypeptides of 48, 84, 91, and 113 kD bound to the ISRE in vitro; the larger three polypeptides most likely compose the ISGF3α component. These ISGF3α polypeptides were unable to bind DNA alone but formed a DNA-binding complex in conjunction with ISGF3γ. The resulting heteromeric complex had the same ISRE-binding specificity as the individual ISGF3γ polypeptide but ~25-fold higher affinity. Whereas ISGF3γ partitioned between the cytoplasm and nucleus in unstimulated cells, ISGF3α was stimulated to translocate to the nucleus only following IFN-α treatment, resulting in preferential nuclear accumulation of both ISGF3α and ISGF3γ as a stable ISGF3-ISRE complex. This regulated nuclear translocation of an activated transcription factor subunit maintained the specificity and rapidity of the IFN-α signaling pathway.

Original languageEnglish (US)
Pages (from-to)1753-1765
Number of pages13
JournalGenes and Development
Volume4
Issue number10
DOIs
StatePublished - Jan 1 1990

Keywords

  • Signal transduction
  • interferon-stimulated genes
  • nuclear translocation
  • transcription
  • transcription factor complex

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology

Fingerprint Dive into the research topics of 'Interferon-α regulates nuclear translocation and DNA-binding affinity of ISGF3, a multimeric transcriptional activator'. Together they form a unique fingerprint.

  • Cite this