The Interferon (IFN)-stimulated Gene Sp100 Promoter Contains an IFN-γ Activation Site and an Imperfect IFN-stimulated Response Element Which Mediate Type I IFN Inducibility

doi:10.1074/jbc.271.41.25253

The Interferon (IFN)-stimulated Gene Sp100 Promoter Contains an IFN-γ Activation Site and an Imperfect IFN-stimulated Response Element Which Mediate Type I IFN Inducibility*

From the Heinrich-Pette-Institut für experimentelle Virologie und Immunologie an der Universität Hamburg, 20251 Hamburg, Germany

^‡ To whom correspondence should be addressed:
Heinrich-Pette-Institut, Martinistr. 52, 20251 Hamburg, Germany
. Tel. and Fax: 49-40-48051221.

Abstract

Expression of the nuclear domain-associated proteins Sp100, PML, and NDP52, is enhanced by interferons (IFNs) on the mRNA and protein level. Increase both of Sp100 and PML mRNA is due to enhanced transcription of the corresponding genes which occurs independently of cellular protein synthesis immediately upon IFN-β addition. Here, we describe the molecular cloning and functional analysis of the Sp100 promoter. DNA sequence analysis revealed potential binding sites for several constitutive and IFN-inducible transcription factors. Consistent with the absence of a TATA box and an initiator element, several transcription initiation sites were found. Transient expression studies identified an imperfect IFN-stimulated response element within the first 100 nucleotides upstream of the major transcription start site. This element rendered a heterologous promoter IFN-β-inducible and bound IFN-stimulated gene factor 2 strongly but IFN-stimulated gene factor 3 only weakly. An IFN-γ activation site approximately 500 base pairs upstream of the IFN-stimulated response element was found to bind three IFN-α/β activation factors upon IFN-β induction and conferred both type I and type II IFN inducibility upon a heterologous promoter. These data demonstrate a novel arrangement of a nonoverlapping IFN-γ activation site and an IFN-stimulated response element mediating type I IFN inducibility, previously not reported for other IFN-stimulable promoters.

Footnotes

↵* This work was supported by a grant from the Bundesministerium für Forschung und Technologie. The Heinrich-Pette-Institut is supported by the Bundesministerium für Gesundheit and the Freie und Hansestadt Hamburg. The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) X95472[GenBank].
↵¹

ND

nuclear dot

IFN

interferon

APL

acute promyelocytic leukemia

STAT

signal transducer and activator of transcription

ISRE

IFN-stimulated response element

ISGF

IFN-stimulated gene factor

IRF

IFN regulatory factor

GAF

IFN-γ activation factor

GAS

IFN-γ activation site

AAF

IFN-α activation factor

HSV

herpes simplex virus

HIP1

housekeeping initiator protein 1

PCR

polymerase chain reaction

CAT

chloramphenicol acetyltransferase

ELISA

enzyme-linked immunosorbent assay

bp

base pair(s).
↵² C. Szostecki, H. H. Guldner, and H. Will, unpublished data.
- Received February 5, 1996.
- Revision received June 20, 1996.