A genome-wide RNAi screen reveals multiple regulators of caspase activation

J Cell Biol. 2007 Nov 19;179(4):619-26. doi: 10.1083/jcb.200708090. Epub 2007 Nov 12.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acetyltransferases / metabolism
  • Animals
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Caspases / analysis
  • Caspases / metabolism*
  • Caspases / physiology*
  • Cell Death / drug effects
  • Cell Death / genetics
  • Cell Survival / drug effects
  • Cells, Cultured
  • DNA Damage
  • Doxorubicin / pharmacology
  • Drosophila Proteins / physiology
  • Drosophila melanogaster / cytology
  • Drosophila melanogaster / embryology
  • Drosophila melanogaster / metabolism
  • Embryo, Nonmammalian
  • Enzyme Activation
  • Epistasis, Genetic
  • Gene Silencing
  • Genome*
  • HeLa Cells
  • Hemocytes / cytology
  • Hemocytes / drug effects
  • Humans
  • Inhibitor of Apoptosis Proteins / physiology
  • N-Terminal Acetyltransferase A
  • N-Terminal Acetyltransferase E
  • Protein-Serine-Threonine Kinases / metabolism
  • RNA Interference*
  • RNA, Small Interfering / metabolism
  • Transcription Factors / physiology
  • Transfection
  • bcl-2 Homologous Antagonist-Killer Protein / metabolism
  • bcl-2-Associated X Protein / metabolism

Substances

  • BAK1 protein, human
  • BAX protein, human
  • Drosophila Proteins
  • Inhibitor of Apoptosis Proteins
  • RNA, Small Interfering
  • Transcription Factors
  • bcl-2 Homologous Antagonist-Killer Protein
  • bcl-2-Associated X Protein
  • chn protein, Drosophila
  • Doxorubicin
  • Acetyltransferases
  • N-Terminal Acetyltransferase A
  • NAA10 protein, human
  • N-Terminal Acetyltransferase E
  • PLK3 protein, human
  • Protein-Serine-Threonine Kinases
  • Caspases