Comparison of the modes of action of a Vero toxin (a Shiga-like toxin) from Escherichia coli, of ricin, and of alpha-sarcin.
The modes of action of a Vero toxin (VT2 or Shiga-like toxin II) from Escherichia coli, of ricin, and of alpha-sarcin were compared. Elongation factor 1 (EF1) and GTP-dependent Phe-tRNA binding to ribosomes in the presence of poly(U) was inhibited by these three toxins, but EF1 and guanylyl (beta, gamma-methylene)-diphosphate-dependent Phe-tRNA binding was inhibited by alpha-sarcin only. EF1- and Phe-tRNA-dependent GTPase activity was inhibited by these toxins, but nonenzymatic binding of Phe-tRNA was not. The turnover rate of EF1 binding to ribosomes during Phe-tRNA binding was also decreased by these three toxins. The addition of EF1 recovered the inhibition of Phe-tRNA binding to ribosomes by VT2 and ricin but not by alpha-sarcin. The formation of and EF2- and GTP-dependent puromycin derivative of phenylalanine was inhibited slightly by the three toxins, indicating that translocation is not influenced significantly by them. EF2-dependent GTPase activity was stimulated by these toxins, and especially by VT2 and ricin. In contrast, the binding of EF2 to ribosomes was inhibited strongly by VT2 and ricin, and slightly by alpha-sarcin. The stimulation of EF2-dependent GTPase activity by the toxins may compensate for the decrease of EF2 binding to ribosomes which they caused during translocation. In total, these results indicate that VT2 and ricin inhibit protein synthesis through the disturbance of the turnover of EF1 binding to ribosomes during aminoacyl-tRNA binding to ribosomes, and that alpha-sarcin inhibits the synthesis through the inhibition of the binding of the complex of Phe-tRNA, EF1, and GTP to ribosomes.
Furutani
M
M
Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Kashiwagi
K
K
Ito
K
K
Endo
Y
Y
Igarashi
K
K
D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology*; D004722:Endoribonucleases; D005656:Fungal Proteins / Q000494:pharmacology*; D005914:Globins / Q000096:biosynthesis; D006160:Guanosine Triphosphate / Q000378:metabolism; D066298:In Vitro Techniques; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects*; D020648:Peptide Elongation Factor 1; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D011817:Rabbits; D012270:Ribosomes / Q000378:metabolism*; D012276:Ricin / Q000633:toxicity*; D022622:Shiga Toxin 1
D001427:Bacterial Toxins; D005656:Fungal Proteins; D020648:Peptide Elongation Factor 1; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors; D012346:RNA, Transfer, Amino Acyl; D022622:Shiga Toxin 1; C014801:alpha-sarcin; D006160:Guanosine Triphosphate; D005914:Globins; D012276:Ricin; D004722:Endoribonucleases