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Research Article

Interferon induction of gene transcription analyzed by in vivo footprinting.

J Mirkovitch, T Decker, J E Darnell Jr
J Mirkovitch
Rockefeller University, New York, New York 10021.
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T Decker
Rockefeller University, New York, New York 10021.
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J E Darnell Jr
Rockefeller University, New York, New York 10021.
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DOI: 10.1128/MCB.12.1.1
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ABSTRACT

The promoters of two interferon-induced genes (the ISG54 and guanylate-binding protein [GBP] genes) have been analyzed in whole cells and in isolated nuclei by using a new genomic sequencing technique. The ISG54 gene contains an interferon-simulating response element (ISRE), earlier shown to be necessary and sufficient for alpha interferon (IFN-alpha) induction, that appeared complexed with proteins in both transcribing and nontranscribing cells. However, the extent of protection and hypersensitivity to DNase I or dimethyl sulfate within the ISRE region was changed upon transcriptional induction, suggesting the binding of different factors in different transcriptional states. In addition to the ISRE, the GBP gene needs a newly recognized DNA element, called the GAS, that partly overlaps the ISRE for full induction by either IFN-alpha or IFN-gamma. This GAS element was transiently protected against DNase I in the nuclei of interferon-treated cells but was not protected at later times when transcription reached maximal levels. Thus, the GAS-binding activity may be necessary only transiently for the initial assembly of a transcription initiation complex on the GBP promoter. Dimethyl sulfate methylation of genomic DNA performed on intact cells showed a characteristic sensitivity over the GAS that correlated with transcription levels and that persisted longer than did DNase I protection over the GAS. These results demonstrate the involvement of the GAS in IFN-alpha and -gamma induction of GBP and suggest the presence of an altered DNA conformation or a small protein in the major groove of the GAS associated with ongoing GBP transcription.

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Interferon induction of gene transcription analyzed by in vivo footprinting.
J Mirkovitch, T Decker, J E Darnell Jr
Molecular and Cellular Biology Jan 1992, 12 (1) 1-9; DOI: 10.1128/MCB.12.1.1

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Interferon induction of gene transcription analyzed by in vivo footprinting.
J Mirkovitch, T Decker, J E Darnell Jr
Molecular and Cellular Biology Jan 1992, 12 (1) 1-9; DOI: 10.1128/MCB.12.1.1
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