Housekeeping genes commanded to commit suicide in CpG-cleavage commitment upstream of Bcl-2 inhibition in caspase-dependent and -independent pathways.
A CpG-specific commitment common to both caspase-dependent and -independent cell deaths implies critical gene activity from epigenetic modulation. Using a focused microarray (genechip) of 22 housekeeping genes, which have canonical CpG islands at 5'-promoter regions, here we show critical regulation of vital intermediary metabolism and cell structure that are common to both caspase-dependent fasL-mediated and caspase-independent etoposide-mediated cell deaths. Gene activity of at least twofold under or over control levels and common to both cell death pathways was considered to be significantly regulated in common. Seven genes controlling energy production in glycolysis, tricarboxylic acid cycle, and the respiratory electron transport chain were significantly downregulated in common. Energy depletion is lethal. Downregulated pyruvate dehydrogenase E1 gene, in addition, suggested primary metabolic acidification. Cell acidification is also lethal. Critical derangement of the cell structure was suggested by common downregulation of the basal histone gene H2A.X which is required for nucleosome assembly. Common upregulation of the alpha-tubulin gene suggested perturbation of vital microtubular dynamics. Gene-commanded cell suicide was suggested. We further show that a Bcl-2 overexpression of three- to fourfold above normal levels could not prevent the CpG-specific megabase DNA cleavages in the two cell death pathways, but abolished their low-molecular-weight 200-bp ladder cleavages. Together with incomplete suppression of the other apoptotic expressions, the Bcl-2 inhibition point appeared downstream from the CpG-cleavage commitment point.
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Department of Anatomy, Faculty of Medicine, National University of Singapore, Kent Ridge, 117597, Singapore.
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D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D020169:Caspases / Q000235:genetics / Q000378:metabolism*; D002453:Cell Cycle; D016923:Cell Death / Q000235:genetics*; D002460:Cell Line; D018899:CpG Islands / Q000235:genetics*; D053938:DNA Fragmentation; D016521:Electrophoresis, Gel, Pulsed-Field; D004734:Energy Metabolism / Q000235:genetics; D005047:Etoposide / Q000494:pharmacology; D053222:Fas Ligand Protein; D005786:Gene Expression Regulation; D019254:Genes, bcl-2 / Q000235:genetics*; D006801:Humans; D008562:Membrane Glycoproteins / Q000494:pharmacology; D020411:Oligonucleotide Array Sequence Analysis; D014407:Tumor Cells, Cultured
D017304:Annexin A5; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D005047:Etoposide; D020169:Caspases