Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1968 Jul;96(1):61–69. doi: 10.1128/jb.96.1.61-69.1968

Response of Cultured Mammalian Cells to Diphtheria Toxin III. Inhibition of Protein Synthesis Studied at the Subcellular Level

Thomas J Moehring a,1, Joan M Moehring a,1
PMCID: PMC252253  PMID: 5663574

Abstract

Diphtheria toxin inhibited protein synthesis in intact KB cells. The action of the toxin upon the cell did not result in disaggregation of polyribosomes, or in impairment of their ability to function in protein synthesis. A reduction in single ribosomes and a concomitant increase in polyribosomes did result from the action of toxin. Nascent peptides were not cleaved from polyribosomes by the action of toxin, but treatment of fully intoxicated cells with puromycin resulted in cleavage of these peptides, and caused accelerated polyribosome breakdown. Our data indicated that the toxin must enter the cell to exert its effect. The component or components sensitive to toxin were localized in the 100,000 × g supernatant fraction of cytoplasmic extracts. When extracts from intoxicated cells were treated with nicotinamide, a significant proportion of their capacity to synthesize protein was restored. The specificity of this reaction suggested that nicotinamide adenine dinucleotide is involved in the action of toxin in the intact cell, and that one component inactivated by toxin is soluble transferase II.

Full text

PDF
61

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ARLINGHAUS R., SHAEFER J., SCHWEET R. MECHANISM OF PEPTIDE BOND FORMATION IN POLYPEPTIDE SYNTHESIS. Proc Natl Acad Sci U S A. 1964 Jun;51:1291–1299. doi: 10.1073/pnas.51.6.1291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonventre P. F., Imhoff J. G. Studies on the mode of action of diphtheria toxin. II. Protein synthesis in primary heart cell cultures. J Exp Med. 1967 Dec 1;126(6):1079–1086. doi: 10.1084/jem.126.6.1079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. COLLIER R. J., PAPPENHEIMER A. M., Jr STUDIES ON THE MODE OF ACTION OF DIPHTHERIA TOXIN. II. EFFECT OF TOXIN ON AMINO ACID INCORPORATION IN CELL-FREE SYSTEMS. J Exp Med. 1964 Dec 1;120:1019–1039. doi: 10.1084/jem.120.6.1019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Collier R. J. Effect of diphtheria toxin on protein synthesis: inactivation of one of the transfer factors. J Mol Biol. 1967 Apr 14;25(1):83–98. doi: 10.1016/0022-2836(67)90280-x. [DOI] [PubMed] [Google Scholar]
  5. Duncan J. L., Groman N. B. Studies of the activity of diphtheria toxin. I. Poliovirus replication in intoxicated HeLa cells. J Exp Med. 1967 Mar 1;125(3):489–500. doi: 10.1084/jem.125.3.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goor R. S., Pappenheimer A. M., Jr, Ames E. Studies on the mode of action of diphtheria toxin. V. Inhibition of peptide bond formation by toxin and NAD in cell-free systems and its reversal by nicotinamide. J Exp Med. 1967 Nov 1;126(5):923–939. doi: 10.1084/jem.126.5.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goor R. S., Pappenheimer A. M., Jr Studies on the mode of action of diphtheria toxin. 3. Site of toxin action in cell-free extracts. J Exp Med. 1967 Nov 1;126(5):899–912. doi: 10.1084/jem.126.5.899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goor R. S., Pappenheimer A. M., Jr Studies on the mode of action of diphtheria toxin. IV. Specificity of the cofactor (NAD) requirement for toxin action in cell-free systems. J Exp Med. 1967 Nov 1;126(5):913–921. doi: 10.1084/jem.126.5.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. KATO I., PAPPENHEIMER A. M., Jr An early effect of diphtheria toxin on the metabolism of mammalian cells growing in culture. J Exp Med. 1960 Aug 1;112:329–349. doi: 10.1084/jem.112.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Moehring J. M., Moehring T. J. The response of cultured mammalian cells to diphtheria toxin. II. The resistant cell: enhancement of toxin action by poly-L-ornithine. J Exp Med. 1968 Mar 1;127(3):541–554. doi: 10.1084/jem.127.3.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Moehring T. J., Moehring J. M., Kuchler R. J., Solotorovsky M. The response of cultured mammalian cells to diphtheria toxin. I. Amino acid transport, accumulation, and incorporation in normal and intoxicated sensitive cells. J Exp Med. 1967 Sep 1;126(3):407–422. doi: 10.1084/jem.126.3.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Monro R. E., Marcker K. A. Ribosome-catalysed reaction of puromycin with a formylmethionine-containing oligonucleotide. J Mol Biol. 1967 Apr 28;25(2):347–350. doi: 10.1016/0022-2836(67)90146-5. [DOI] [PubMed] [Google Scholar]
  13. STRAUSS N., HENDEE E. D. The effect of diphtheria toxin on the metabolism of HeLa cells. J Exp Med. 1959 Feb 1;109(2):145–163. doi: 10.1084/jem.109.2.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. TRAUT R. R., MONRO R. E. THE PUROMYCIN REACTION AND ITS RELATION TO PROTEIN SYNTHESIS. J Mol Biol. 1964 Oct;10:63–72. doi: 10.1016/s0022-2836(64)80028-0. [DOI] [PubMed] [Google Scholar]
  15. WILLIAMSON A. R., SCHWEET R. ROLE OF THE GENETIC MESSAGE IN INITIATION AND RELEASE OF THE POLYPEPTIDE CHAIN. Nature. 1964 May 2;202:435–437. doi: 10.1038/202435a0. [DOI] [PubMed] [Google Scholar]
  16. WILLIAMSON A. R., SCHWEET R. ROLE OF THE GENETIC MESSAGE IN POLYRIBOSOME FUNCTION. J Mol Biol. 1965 Feb;11:358–372. doi: 10.1016/s0022-2836(65)80063-8. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES