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Activation of phosphoinositide 3-kinase by the NBS1 DNA repair protein through a novel activation motif

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Abstract

Class IA phosphoinositide 3-kinases (PI 3-kinases) are key signaling components downstream of tyrosine kinases and Ras, regulating many different cellular functions and contributing to tumorigenesis. Class IA PI 3-kinases are heterodimers comprised of a p85 regulatory and a p110 catalytic subunit. Nijmegen breakage syndrome (NBS) is a chromosomal instability syndrome associated with cancer predisposition, radiosensitivity, microcephaly, and growth retardation. The NBS gene product p95 (also known as NBS1) is part of the Mre11-Rad50-Nbs1 complex, a central player associated with double-strand break repair. We previously demonstrated that NBS1 overexpression induces transformation through activation of PI 3-kinase/Akt. In this study, we show that NBS1 directly interacts, through a highly conserved C-terminal motif (aa 653–669) of NBS1, with the N-terminal domain (aa 1–108) of the p110α catalytic subunit of PI 3-kinase, and stimulates PI 3-kinase activity. Mutations of different regions of the conserved motif abolish the ability of NBS1 to activate PI 3-kinase in vitro and in vivo. Co-expression of NBS1/p110α/p-Akt is observed in certain percentage of head and neck cancer patient samples. These results demonstrate that NBS1 can function as an adaptor/activator of p110α PI 3-kinase through a novel activation motif, consistent with its possible role in cell transformation and tumorigenesis.

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Abbreviations

NBS:

Nijmegen breakage syndrome

PI 3-kinase:

phosphoinositide 3-kinase

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Acknowledgments

We thank Drs. Y.C.Wu, J.H.Lin, A.M.Lin, and the Genomic Center of National Yang-Ming University for providing pGEX-4T−1, mouse p110α, pFlag-CMV2, and human p110γ cDNA clones, respectively. We are grateful to the excellent technical assistance of W.C. Chiang. We appreciate Drs. T.Y. Chou, W.Y. Li, and C.J. Liu (Taipei Mackay Memorial Hospital) for their reading of the IHC results and providing patient samples. This work was supported in part by National Research Program for Genomic Medicine (DOH-96-TD-G-111-002) (K.J.W.), Taipei Veterans General Hospital V96-ER2-010 (S.Y.C), VGH 96-C1-126, V-96-ER2-008 (M.H.Y.), National Science Council (NSC-95-2320-B-010-065) (K.J.W.), a grant from Ministry of Education, Aim for the Top University Plan (96A-D-D139)(K.J.W.), and National Health Research Institutes (NHRI-EX96-9611BI) (K.J.W.).

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Authors and Affiliations

  1. Institute of Biochemistry and Molecular Biology, National Yang-Ming University, No.155, Li-Nong St., Sec.2, Peitou, Taipei, 112, Taiwan

    Yen-Chung Chen, Hsiu-Yin Chiang & Kou-Juey Wu

  2. Clinical Medicine, National Yang-Ming University, Taipei, 112, Taiwan

    Muh-Hwa Yang & Kou-Juey Wu

  3. Division of Oncology, Departments of Medicine, Taipei Veterans’ General Hospital, Taipei, 112, Taiwan

    Muh-Hwa Yang & Po-Min Chen

  4. Otolaryngology, Taipei Veterans’ General Hospital, Taipei, 112, Taiwan

    Shyue-Yih Chang

  5. Genomic Research Center, Taipei Veterans’ General Hospital, Taipei, 112, Taiwan

    Muh-Hwa Yang, Po-Min Chen & Kou-Juey Wu

  6. Graduate Institute of Microbiology, College of Medicine, National Taiwan University, Taipei, 100, Taiwan

    Shu-Chun Teng

  7. Ludwig Institute for Cancer Research, 91 Riding House Street, London, W1W 7BS, UK

    Bart Vanhaesebroeck

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  1. Yen-Chung Chen
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Correspondence to Kou-Juey Wu.

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Supplementary Table 1

Sequence of the oligonucleotides and restriction enzymes used for plasmid construction (DOC 190 KB)

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Chen, YC., Chiang, HY., Yang, MH. et al. Activation of phosphoinositide 3-kinase by the NBS1 DNA repair protein through a novel activation motif. J Mol Med 86, 401–412 (2008). https://doi.org/10.1007/s00109-008-0302-x

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  • DOI: https://doi.org/10.1007/s00109-008-0302-x

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