Identification of GRIM-19, a novel cell death-regulatory gene induced by the interferon-beta and retinoic acid combination, using a genetic approach.
We show here that the combination of interferon-beta (IFN-beta) 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-IFN-induced 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. Antisense expression of GRIM-19 confers a strong resistance against IFN/RA-induced death by reducing the intracellular levels of GRIM-19 protein. Overexpression of GRIM-19 enhances cell death in response to IFN/RA. GRIM-19 is primarily a nuclear protein whose expression is induced by the IFN/RA combination. Together, our studies identify a novel cell death-regulatory molecule.
Angell
J E
JE
Greenebaum Cancer Center, Department of Microbiology and Immunology, Molecular and Cellular Biology Program, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA.
Lindner
D J
DJ
Shapiro
P S
PS
Hofmann
E R
ER
Kalvakolanu
D V
DV
D000595:Amino Acid Sequence; D017209:Apoptosis; D001943:Breast Neoplasms / Q000473:pathology; D016373:DNA, Antisense / Q000494:pharmacology; D029721:Drosophila Proteins; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D016899:Interferon-beta / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D013329:Structure-Activity Relationship; D014212:Tretinoin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D016373:DNA, Antisense; D029721:Drosophila Proteins; D009479:Neuropeptides; D012333:RNA, Messenger; C108881:grim protein, Drosophila; D014212:Tretinoin; D016899:Interferon-beta