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Nucleic Acids Research, 1990, Vol. 18, No. 8 2125-2131
© 1990

MOLECULAR BIOLOGY

DNA-protein interactions at the interferon-responsive promoter: evidences for an involvement of phosphorylation

Christian Roy and Bernard Lebleu

UA CNRS 1191, Laboratoire de Biochimie des Protéines, Université de Montpellier II Sciences et Techniques du Languedoc, 34095 Montpellier Cedex 2, France

Received November 23, 1989. Revised January 17, 1990. Accepted January 17, 1990.

Several {alpha}ß and {gamma} interferons (IFN) induced genes are regulated transcriptionally via a 29 bp cis acting regulatory element (ICS, Interferon Consensus Sequence). The ICS binding capacity of HeLa cell nuclear extracts was strictly dependent on the presence of nucleoside triphosphate and Mg2+. It increased upon {alpha}ß or {gamma} IFN treatment of the cells. Three 50 mer synthetic oligonucleotides containing the ICS or putative ISRE (Interferon Stimulatable Response Element), representing portions of genes inducible by {alpha}ß IFN (HLA-A3 and IFI-56K) or by {gamma} (HLA-DR) were used as probes to titrate nuclear factors interacting with the ICS. All three probes were retarded in a mobility shift assay in two bands. Phosphorylation conditions were crucial for demonstrating their existence and/or their relative amounts. A factor whose activity and/or amount was modulated upon IFN treatment, appeared to be involved in phosphorylation dependent posttransiational modification(s) of the ISRE binding proteins responsible for altered binding properties.


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