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Nuclear import and subnuclear localization of the proto-oncoprotein ETO (MTG8)

Oncogene volume 19, pages 3584–3597 (2000)Cite this article

Abstract

ETO (MTG8) was first described due to its involvement in the (8;21) translocation frequently observed in acute myeloid leukemias. In the t(8;21) the AML1 gene on chromosome 21 is fused to ETO on chromosome 8. The resultant hybrid protein is comprised of the DNA binding domain of AML-1 and the majority of ETO. This study examines the subnuclear distributions of ETO, AML-1B and AML-1/ETO proteins fused to green fluorescence protein in living cells using fluorescence microscopy. Further, we identified a 40 amino acid portion of ETO (amino acids 241–280) that was sufficient to cause nuclear import of green fluorescent protein. Mutational analysis demonstrated that lysine 265 and/or arginine 266 were required for nuclear import of ETO, but that the surrounding basic residues were not critical. ETO interacted with the nuclear import proteins importin-α and β in vitro, and mutations in ETO that abolish nuclear localization also abolished the in vitro interaction with importin-α and β. These data suggest that ETO enters the nucleus via an importin-mediated pathway. Additionally, ETO and AML-1/ETO co-localized to punctate nuclear bodies distinct from those containing promyelocytic leukemia protein. Nuclear body formation was dependent upon a region of ETO N-terminal to the nuclear localization signal. Thus, ETO and AML-1/ETO reside in potentially novel subnuclear compartments.

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Acknowledgements

The authors would like to thank Dr K Tatchell for his help and Drs JN Davis and S Kornbluth for the gift of the GFP antiserum and pGEX-importin-α and β vectors, respectively. S Meyers would especially like to thank Ellie Shaina Davis. This work is in partial fulfillment of a master degree for Y Odaka and was supported by a grant from the National Cancer Institute (CA76186) to S Meyers and by funds provided by the Feist-Weiler Cancer Center and the Virginia K Shehee Biomedical Research Foundation.

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  1. Department of Biochemistry and Molecular Biology, Louisiana State University Health Science Center School of Medicine, Shreveport, Louisiana, USA

    Yoshinobu Odaka, Anirudh Mally, Liza T Elliott & Shari Meyers

  2. Feist-Weiler Cancer Center, Louisiana State University Health Science Center School of Medicine, Shreveport, Louisiana, USA

    Yoshinobu Odaka, Liza T Elliott & Shari Meyers

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Odaka, Y., Mally, A., Elliott, L. et al. Nuclear import and subnuclear localization of the proto-oncoprotein ETO (MTG8). Oncogene 19, 3584–3597 (2000). https://doi.org/10.1038/sj.onc.1203689

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