Identification of GRIM-19, a Novel Cell Death-regulatory Gene Induced by the Interferon-β and Retinoic Acid Combination, Using a Genetic Approach

Abstract

We show here that the combination of interferon-␤ (IFN-␤) and all-trans-retinoic acid (RA) induces the death of tumor cells. To understand the molecular basis for synergistic growth-suppressive action and to identify the gene products that participate in this process, we have employed an antisense knock-out technique. This approach permits the isolation of cell death-associated genes based on their selective inactivation by overexpression of antisense cDNAs. Because the antisense mRNA inactivates gene expression of death-specific genes, transfected cells survive in the presence death inducers. Several Genes associated with Retinoid-IFNinduced Mortality (GRIM) were identified using this approach. Here we report the isolation of a novel GRIM gene, GRIM-19. This 552-base pair cDNA encodes a 16-kDa protein.

Coupling mitochondrial respiratory chain to cell death: an essential role of mitochondrial complex I in the interferon-β and retinoic acid-induced cancer cell death

“…20,21 It has been shown previously that IFN-b/RA treatment increases the mRNA level of GRIM-19 in human breast carcinoma cell lines. 10 In this study, we investigated whether IFN-b/RA treatment can also stimulate expression of other MRC subunits in these cancer cell lines. We chose certain subunits encoded by the nucleus from complex I-V based on the antibody availability.…”

“…8,9 The discovery of GRIM-19 as an IFN-b/RA-inducible gene product 10 role of MRC in cell death seems specific only for the combination of IFN-b and RA, as IFN or RA alone did not induce either upregulation of the MRC subunits or the cell death (Figures 1 and 2). Furthermore, other death stimuli such as UV, CPT, and staurosporine caused cell death without inducing the expression of GRIM-19 and NDUFS3, and knockdown of these two genes did not prevent cell death induced by these generic death agents (Supplementary Figure 2).…”

“…Other mechanisms such as nuclear translocation were also investigated. GRIM-19 was originally claimed as a nuclear protein, 10 but was later shown to be a mitochondrial protein. 11,12,14 In a recent review by Kalvakolanu, 34 it was proposed that GRIM-19 may initiate apoptosis, like cytochrome c and AIF, by translocation from the mitochondria to the nucleus upon IFN-b/RA stimulation.…”

“…10 GRIM-19 was initially implicated as a nuclear protein, and its precise role in the IFN-b-and RA-induced tumor cell death was not clear. 10 Subsequently, the bovine GRIM-19 homolog was found to copurify with mitochondrial NADH:ubiquinone oxidoreductase (complex I) in the heart. 11 GRIM-19 was also identified as a negative regulator of signal transducer and activator of transcription (Stat)3.…”

Identification of alternatively spliced GRIM-19 mRNA in kidney cancer tissues

“…12 Gene associated with Retinoid-Interferon-induced Mortality (GRIM)-19 was originally defined as a cell death regulatory gene, as it is essential for the tumor cell death induced by interferon-b and retinoic acid. 13 Subsequently, GRIM-19 was demonstrated to be a new subunit of the mitochondrial NAPDH:ubiquinone oxidoredutase (respiratory chain complex I) and is essential for the complex I assembly, activity and maintenance of mitochondrial membrane potential. [14][15][16][17] Disruption of mitochondrial transmembrane potential by GRIM-19 mutants enhances the cells' sensitivity to apoptotic stimuli.…”