Inducible cell ablation in Drosophila by cold-sensitive ricin A chain.
We have developed a system for temperature-inducible killing of specific cells in the fruitfly Drosophila melanogaster. The system overcomes many of the limitations of existing cell ablation methods and is in principle applicable to any non-homeothermic eukaryote. Temperature-sensitive and cold-sensitive mutations in the ricin toxin A chain (RTA) of castor bean were generated in yeast. One cold-sensitive mutation, RAcs2, produced temperature-dependent ablation of eye cells in Drosophila when expressed under control of the eye-specific sev enhancer. At 29 degrees C, cell death was observed within 7 hours in the developing eye and no obvious toxic effects were observed elsewhere; at 18 degrees C, extremely low toxicity was observed. DNA sequencing of RAcs2 revealed a single amino acid substitution in the RTA active site cleft.
Moffat
K G
KG
Department of Biological Sciences, University of Warwick, Coventry, UK.
Gould
J H
JH
Smith
H K
HK
O'Kane
C J
CJ
D000818:Animals; D016923:Cell Death / Q000235:genetics*; D003080:Cold Temperature; D004331:Drosophila melanogaster / Q000235:genetics*; D005123:Eye / Q000166:cytology / Q000196:embryology / Q000648:ultrastructure; D005804:Genes, Lethal / Q000502:physiology*; D008855:Microscopy, Electron, Scanning; D012276:Ricin / Q000235:genetics*
D012276:Ricin