A genome-wide RNAi screen reveals multiple regulators of caspase activation.
Apoptosis is an evolutionally conserved cellular suicide mechanism that can be activated in response to a variety of stressful stimuli. Increasing evidence suggests that apoptotic regulation relies on specialized cell death signaling pathways and also integrates diverse signals from additional regulatory circuits, including those of cellular homeostasis. We present a genome-wide RNA interference screen to systematically identify regulators of apoptosis induced by DNA damage in Drosophila melanogaster cells. We identify 47 double- stranded RNA that target a functionally diverse set of genes, including several with a known function in promoting cell death. Further characterization uncovers 10 genes that influence caspase activation upon the removal of Drosophila inhibitor of apoptosis 1. This set includes the Drosophila initiator caspase Dronc and, surprisingly, several metabolic regulators, a candidate tumor suppressor, Charlatan, and an N-acetyltransferase, ARD1. Importantly, several of these genes show functional conservation in regulating apoptosis in mammalian cells. Our data suggest a previously unappreciated fundamental connection between various cellular processes and caspase-dependent cell death.
Yi
Caroline H
CH
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Sogah
Dodzie K
DK
Boyce
Michael
M
Degterev
Alexei
A
Christofferson
Dana E
DE
Yuan
Junying
J
D000123:Acetyltransferases / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D020169:Caspases / Q000032:analysis / Q000378:metabolism* / Q000502:physiology*; D016923:Cell Death / Q000187:drug effects / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004249:DNA Damage; D004317:Doxorubicin / Q000494:pharmacology; D029721:Drosophila Proteins / Q000502:physiology; D004331:Drosophila melanogaster / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D004625:Embryo, Nonmammalian; D004789:Enzyme Activation; D004843:Epistasis, Genetic; D020868:Gene Silencing; D016678:Genome; D006367:HeLa Cells; D006434:Hemocytes / Q000166:cytology / Q000187:drug effects; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000502:physiology; D063209:N-Terminal Acetyltransferase A; D063213:N-Terminal Acetyltransferase E; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D014157:Transcription Factors / Q000502:physiology; D014162:Transfection; D025521:Tumor Suppressor Proteins; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D000097868:Polo-like Kinases
D000123:Acetyltransferases; D020169:Caspases; D004317:Doxorubicin; D029721:Drosophila Proteins; D051034:Inhibitor of Apoptosis Proteins; D063209:N-Terminal Acetyltransferase A; D063213:N-Terminal Acetyltransferase E; D017346:Protein Serine-Threonine Kinases; D034741:RNA, Small Interfering; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D050998:bcl-2 Homologous Antagonist-Killer Protein; D051028:bcl-2-Associated X Protein; C494772:BAK1 protein, human; C494826:BAX protein, human; C500129:chn protein, Drosophila; C504824:NAA10 protein, human; C486256:PLK3 protein, human; D000097868:Polo-like Kinases