Isolation of temperature-sensitive diphtheria toxins in yeast and their effects on Drosophila cells

Development. 1992 Mar;114(3):787-96.

Abstract

We have isolated temperature-sensitive diphtheria toxins (DT-A(ts)) to develop a method that allows temporal impedement of cellular functions. Four DT-A(ts) genes were isolated in a mutagenesis screen using the yeast, Saccharomyces cerevisiae. When expressed in yeast, these DT-A(ts) arrest growth at 18 degrees C but not at 30 degrees C. Three DT-A(ts) were subsequently tested in the R1-R6 photoreceptor cells of transgenic fruit flies, Drosophila melanogaster. The toxins show similar temperature dependence in both organisms, suggesting that they may be useful in a wide range of non-homeothermic species. DNA sequence analysis revealed that three of the four DT-A(ts) mutations are novel. Interestingly, the fourth DT-A(ts) carries the same point mutation as the extensively characterized CRM197, an ADP ribosyltransferase-defective form of diphtheria toxin.

Publication types

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

MeSH terms

  • Animals
  • Cell Death
  • Diphtheria Toxin / isolation & purification*
  • Drosophila
  • Female
  • Genes, Bacterial / physiology*
  • Male
  • Microscopy, Electron
  • Mutagenesis
  • Photoreceptor Cells / injuries
  • Photoreceptor Cells / ultrastructure
  • Saccharomyces cerevisiae / chemistry*
  • Sequence Alignment
  • Temperature*

Substances

  • Diphtheria Toxin