Isolation of DAP3, a Novel Mediator of Interferon-
-induced Cell Death (*)
- Joseph L. Kissil,
- Louis P. Deiss(§),
- Michael Bayewitch,
- Tal Raveh,
- George Khaspekov(¶) and
- Adi Kimchi(**)
- From the Department of Molecular Genetics and Virology, The Weizmann Institute of Science, Rehovot 76100, Israel
- ** Incumbent of the Helena Rubinstein Chair in Cancer Research. To whom correspondence should be addressed. Tel.: 972-8-342428; Fax: 972-8-344108.
Abstract
Interaction of certain cytokines with their corresponding cell-surface receptors induces programed cell death. Interferon-
induces in HeLa cells a type of cell death with features characteristic of programed cell death. Here, we report the isolation
of a novel gene, DAP3 (death-associated protein-3), involved in mediating interferon-
-induced cell death. The rescue of this gene was performed by a functional selection approach of gene cloning that is based
on transfection with an antisense cDNA expression library. The antisense RNA-mediated inactivation of the DAP3 gene protected the cells from interferon-
-induced cell death. This property endowed the cells expressing it with a growth advantage in an environment restrictive due
to the continuous presence of interferon-
and thus provided the basis of its selection. The gene is transcribed into a single 1.7-kilobase mRNA, which is ubiquitously
expressed in different tissues and codes for a 46-kDa protein carrying a potential P-loop motif. Ectopic expression of DAP3 in HeLa cells was not compatible with cell growth, resulting in a 16-fold reduction in the number of drug-resistant stable
clones. The data presented suggest that DAP3 is a positive mediator of cell death induced by interferon-
.
Footnotes
-
↵* This work was supported by the Israel Academy of Sciences and Humanities. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by 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) X83544[GenBank].
-
↵1 The abbreviations used are:
- IFN-
-
interferon-
- PAGE
-
polyacrylamide gel electrophoresis
- ORF
-
open reading frame.
- IFN-
-
- Received March 31, 1995.
- Revision received August 11, 1995.
- © 1995 by The American Society for Biochemistry and Molecular Biology, Inc.
