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A CRM1-mediated nuclear export signal governs cytoplasmic localization of BRCA2 and is essential for centrosomal localization of BRCA2

Oncogene volume 27, pages 2969–2977 (2008)Cite this article

Abstract

Germ-line mutations of the BRCA2 gene cause inherited susceptibility to breast and ovarian cancers. BRCA2 contains two nuclear localization signals, predominantly localizes in the nucleus and plays significant roles in DNA double-strand break repair. Recently, we reported that BRCA2 localizes to the centrosomes during the S and early M phases of the cell cycle. In this study, for the first time, we identified a functional nuclear export sequence (NES1; 1383DLSDLTFLEVA1393) in BRCA2. The green fluorescent protein (GFP)-NES1 fusion protein was localized in the cytoplasm and could be blocked by the chromosomal region maintenance 1-specific export inhibitor leptomycin B. Mutation of a leucine residue in the NES1 motif to alanine (L1384A) resulted in both cytoplasmic and nuclear localization of the GFP-NES1 fusion protein and a nuclear accumulation of ectopic full-length BRCA2-FLAG. Moreover, treatment of cells with leptomycin B decreased centrosomal localization of BRCA2. Finally, by microinjection of an anti-BRCA2 antibody into the cytoplasm of HeLa S3 cells, we found that depletion of normal BRCA2 proteins in the cytoplasm leads to centrosome amplification and binucleated cells. Our results suggest that disruption of the NES function by genetic changes results in deregulation of BRCA2 export, which ultimately leads to centrosome disorder.

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Acknowledgements

We thank all members of Department of Molecular Diagnosis of the Cancer Institute, Japanese Foundation for Cancer Research for helpful discussions and encouragement. Itaru Sakoh, Koki Miyamoto, Miki Fukuda at the Fujitsu co. are thanked for performing the microinjections. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology in Japan.

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Authors and Affiliations

  1. Department of Molecular Genetics, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan

    X Han & Y Miki

  2. Department of Preventive Medicine, Yanbian University College of Medicine, Yanji, China

    X Han

  3. Department of Genetic Diagnosis, The Cancer Institute, Japanese Foundation for Cancer Research, Tokyo, Japan

    H Saito, Y Miki & A Nakanishi

  4. Institute of Medical Science, Tokyo University, Tokyo, Japan

    A Nakanishi

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  1. X Han
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  2. H Saito
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Correspondence to Y Miki.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Han, X., Saito, H., Miki, Y. et al. A CRM1-mediated nuclear export signal governs cytoplasmic localization of BRCA2 and is essential for centrosomal localization of BRCA2. Oncogene 27, 2969–2977 (2008). https://doi.org/10.1038/sj.onc.1210968

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