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. 2013 Jan;131(1):153-63.
doi: 10.1093/toxsci/kfs266. Epub 2012 Sep 11.

Modulation of inflammatory gene expression by the ribotoxin deoxynivalenol involves coordinate regulation of the transcriptome and translatome

Kaiyu He  1 , Xiao Pan, Hui-Ren Zhou, James J Pestka
Affiliations

Modulation of inflammatory gene expression by the ribotoxin deoxynivalenol involves coordinate regulation of the transcriptome and translatome

Kaiyu He et al. Toxicol Sci. 2013 Jan.

Abstract

The trichothecene deoxynivalenol (DON), a common contaminant of cereal-based foods, is a ribotoxic mycotoxin known to activate innate immune cells in vivo and in vitro. Although it is recognized that DON induces transcription and mRNA stabilization of inflammation-associated mRNAs in mononuclear phagocytes, it is not known if this toxin affects translation of selected mRNA species in the cellular pool. To address this question, we employed a focused inflammation/autoimmunity PCR array to compare DON-induced changes in profiles of polysome-associated mRNA transcripts (translatome) to total cellular mRNA transcripts (transcriptome) in the RAW 264.7 murine macrophage model. Exposure to DON at 250 ng/ml (0.84 µM) for 6 h induced robust expression changes in inflammatory response genes including cytokines, cytokine receptors, chemokines, chemokine receptors, and transcription factors, with 73% of the changes being highly comparable within transcriptome and translatome populations. When expression changes of selected representative inflammatory response genes in the polysome and cellular mRNA pools were quantified in a follow-up study by real-time PCR, closely coordinated regulation of the translatome and transcriptome was confirmed; however, modest but significant differences in the relative expression of some genes within the two pools were also detectable. Taken together, DON's capacity to alter translation expression of inflammation-associated genes appears to be driven predominantly by selective transcription and mRNA stabilization that have been reported previously; however, a small subset of these genes appear to be further regulated at the translational level.

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Figures

FIG. 1.
FIG. 1.
Relative numbers of array genes by DON in the transcriptome and translatome. Based on the PCR array result, the percentage of DON-induced up-, down-, and unregulated genes were calculated and shown in (A) transcriptome and (B) translatome, respectively.
FIG. 2.
FIG. 2.
Comparison of DON overlapping genes in transcriptome and translatome. Numbers of genes (A) upregulated and (B) downregulated by DON in transcriptome and translatome, respectively. Overlapping regions represent the common genes that were shared by transcriptome and translatome.
FIG. 3.
FIG. 3.
Scatter distribution of up- and downregulated genes in transcriptome and translatome. (A) Transcriptome and (B) translatome data (DON vs. Control) were plotted using SAbiosciences web-based RT2 Profiler PCR Array Data Analysis tool and exported. Each dot represents a single gene. The parallel line region indicates twofold threshold and the black arrows demonstrate the up- or downregulation of genes. Examples of commonly upregulated (CCL3, CCL4, CXCL2, CCR1, CCR2, CCR3, and CCL7) and downregulated genes (LTB, IL-7, IL-18, CXCL10, and CD40) are identified.
FIG. 4.
FIG. 4.
Comparative effects of DON on cytokine mRNA expression in transcriptome and translatome. Three independent cell culture experiments were conducted and transcriptome and translatome were analyzed by real-time PCR in duplicate. The relative changes in mRNA reflect the ratio of response by DON- and vehicle-treated cells. Data are mean ± SE of triplicate wells. The dotted line indicates the basal level of gene expression (onefold) in total and polysome controls. Asterisk indicates induced significant increases in mRNA expression relative to respective controls within the transcriptome and translatome (p < 0.05).
FIG. 5.
FIG. 5.
Comparative effects of DON on chemokines and their receptors expression in transcriptome and translatome. Study was conducted and analyzed as described in the legend of Figure 4.
FIG. 6.
FIG. 6.
Comparative effects of DON on transcription factor mRNA expression in transcriptome and translatome. Study was conducted and analyzed as described in the legend of Figure 4.
FIG. 7.
FIG. 7.
DON induces translatome-specific mRNA expression. Study was conducted and analyzed as described in the legend of Figure 4.
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