ToxSci Advance Access published online on June 4, 2008
Toxicological Sciences, doi:10.1093/toxsci/kfn111
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Comparative Induction of 28S Ribosomal RNA Cleavage by Ricin and the Trichothecenes Deoxynivalenol and T-2 Toxin in the Macrophage
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* Department of Microbiology and Molecular Genetics
Food Science and Human Nutrition
Center for Integrative Toxicology, Michigan State University, East Lansing, MI 48824
1 To whom correspondence should be addressed at 234 G.M. Trout Building, Michigan State University, East Lansing, MI 48824-1224. Fax: (517) 353-8963. E-mail: pestka{at}msu.edu.
Received April 22, 2008; revision received May 29, 2008; accepted May 30, 2008
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Abstract |
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Ribosome inactivating proteins (RIPs) and sesquiterpenoid trichothecene mycotoxins are known to bind to eukaryotic ribosomes, inhibit translation and activate mitogen-activated protein kinases. Here we compared the capacities of the RIP ricin to promote 28S ribosomal RNA (rRNA) cleavage to that of the trichothecenes, deoxynivalenol (DON) and T-2 toxin (T-2). In a cell-free model, exposure to ricin at 300 ng/ml for 30 min depurinated yeast 28S rRNA, however, neither DON (
4 µg/ml) nor T-2 ( 2 µg/ml) exhibited this N-glycosidase activity. Incubation of RAW 264.7 macrophages with ricin (20 to 320 ng/ml), DON (250 to 5000 ng/ml) or T-2 (2 to 80 ng/ml) for 6 h, however, generated 28S rRNA-specific products consistent with cleavage sites near the 3’ terminal end of murine 28S rRNA. Oligonucleotide extension analysis of treated RAW 264.7 cells revealed that ricin evoked 28S rRNA damage at one site in the 
-sarcin/ricin(S/R)-loop (A4256) and two other sites (A3560 and A4045) in the peptidyl transferase center. Although DON or T-2 did not damage the S/R loop, these trichothecenes did promote cleavage at A3560 and A4045. In addition, incubation of the cells with ricin (
20 ng/ml), DON (250 ng/ml) or T-2 (10ng/ml) induced RNase activity as well as RNase L mRNA and protein expression These data suggest that only ricin directly damaged 28S rRNA under cell-free conditions but that ricin, DON and T-2 promoted intracellular 28S rRNA cleavage, potentially by facilitating the action of endogenous RNases and/or by upregulating RNase expression.