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Novel role of hnRNP-A2/B1 in modulating aryl hydrocarbon receptor ligand sensitivity

  • Molecular Toxicology
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Abstract

The aryl hydrocarbon receptor (AHR) is responsible for susceptibility to its ligand-dependent responses. However, the effect of non-AHR factors is less clear. To explore the non-AHR factors, we used two mouse strains with different AHR genetic variants, namely C3H/lpr and MRL/lpr strains with Ala and Val as the 375th amino acid residue, respectively. To assess the contribution of AHR alone, COS-7 cells transiently expressing AHR from each strain were treated with 6-formylindolo[3,2-b]carbazole (FICZ) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and xenobiotic-responsive element (XRE)-driven reporter gene activities were measured. FICZ-EC50 values for the C3H/lpr and MRL/lpr AHR-mediated transactivation were 0.023 and 0.046 nM, respectively, indicating a similar susceptibility in both AHR genotypes. In contrast, C3H/lpr AHR was fourfold more sensitive to TCDD than MRL/lpr AHR. By a pull-down assay using a XRE-containing PCR product as bait and the hepatic nuclear extracts of both FICZ-treated mouse strains, we identified two interacting proteins as heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP-A2) and its splicing variant (hnRNP-A2b). Immunoprecipitation assays demonstrated the AHR interaction with hnRNP-A2/B1. When hnRNP-A2 was co-expressed with the MRL/lpr or C3H/lpr AHR in COS-7, FICZ treatment decreased EC50 to about threefold in both AHR genotypes, compared with EC50 in AHR alone. Similarly, hnRNP-A2b co-expression also lowered the FICZ-EC50 values. In TCDD-treated COS-7, responses depended on the AHR genotype; while no change in TCDD-EC50 was observed for C3H/lpr AHR when hnRNP-A2 was co-expressed, the value was reduced to nearly tenfold for MRL/lpr AHR. Co-transfection with hnRNP-A2b attenuated the AHR sensitivity to TCDD. In conclusion, the hnRNP-A2/B1 interacting with AHR may be a modulator of the AHR ligand sensitivity.

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Acknowledgments

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology to E.-Y. Kim (2013R1A1A2A10010043 and 2012K2A2A4021504). This study was also supported by Grants-in-Aid for Scientific Research (S) (Nos. 21221004 and 26220103) and Challenging Exploratory Research (No. 25660228) to H.I. from Japan Society for the Promotion of Science.

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  1. Masato Nose

    Present address: Department of Histopathology, Tohoku University Graduate School of Medicine, Sendai, Japan

Authors and Affiliations

  1. Department of Life and Nanopharmaceutical Science, Kyung Hee University, Hoegi Dong, Dongdaemun-Gu, Seoul, 130-701, Korea

    See-Wun Cho & Eun-Young Kim

  2. Department of Biology, Kyung Hee University, Hoegi Dong, Dongdaemun-Gu, Seoul, 130-701, Korea

    See-Wun Cho & Eun-Young Kim

  3. Graduate School of Science, Hiroshima University, Hiroshima, Japan

    Ken-ichi Suzuki

  4. Center for Marine Environmental Studies (CMES), Ehime University, Matsuyama, Japan

    Yoshiaki Miura & Hisato Iwata

  5. Department of Pathogenomics, Ehime University Graduate School of Medicine, Toon, Japan

    Tatsuhiko Miyazaki & Masato Nose

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  1. See-Wun Cho
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Cho, SW., Suzuki, Ki., Miura, Y. et al. Novel role of hnRNP-A2/B1 in modulating aryl hydrocarbon receptor ligand sensitivity. Arch Toxicol 89, 2027–2038 (2015). https://doi.org/10.1007/s00204-014-1352-1

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