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Random mutagenesis of PDZOmi domain and selection of mutants that specifically bind the Myc proto-oncogene and induce apoptosis

Oncogene volume 22, pages 2772–2781 (2003)Cite this article

Abstract

Omi is a mammalian serine protease that is localized in the mitochondria and released to the cytoplasm in response to apoptotic stimuli. Omi induces cell death in a caspase-dependent manner by interacting with the X-chromosome linked inhibitor of apoptosis protein, as well as in a caspase-independent way that relies on its proteolytic activity. Omi is synthesized as a precursor polypeptide and is processed to an active serine protease with a unique PDZ domain. PDZ domains recognize the extreme carboxyl terminus of target proteins. Internal peptides that are able to fold into a β-finger are also reported to bind some PDZ domains. Using a modified yeast two-hybrid system, PDZOmi mutants were isolated by their ability to bind the carboxyl terminus of human Myc oncoprotein in yeast as well as in mammalian cells. One such PDZm domain (PDZ-M1), when transfected into mammalian cells, was able to bind to endogenous Myc protein and induce cell death. PDZ-M1-induced apoptosis was entirely dependent on the presence of Myc protein and was not observed when c-myc null fibroblasts were used. Our studies indicate that the PDZ domain of Omi can provide a prototype that could easily be exploited to target specifically and inactivate oncogenes by binding to their unique carboxyl terminus.

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Acknowledgements

We are grateful to members of the Zervos' lab and L Von Kalm for comments, suggestions and critical reading of the manuscript. This work was supported by the National Institutes of Health Grant R01 DK55734-01 (to ASZ).

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Author notes

  1. Daniela Junqueira and Lucia Cilenti: These authors contributed equally to the work

Authors and Affiliations

  1. Biomolecular Science Center and Department of Molecular Biology and Microbiology, University of Central Florida (UCF), 12722 Research Parkway, Orlando, 32826, FL, USA

    Daniela Junqueira, Lucia Cilenti, Lucia Musumeci & Antonis S Zervos

  2. Department of Molecular Biology, Cell Biology, and Biochemistry, Brown University, 69 Brown Street, Providence, 02912, RI, USA

    John M Sedivy

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  1. Daniela Junqueira
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  2. Lucia Cilenti
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Correspondence to Antonis S Zervos.

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Junqueira, D., Cilenti, L., Musumeci, L. et al. Random mutagenesis of PDZOmi domain and selection of mutants that specifically bind the Myc proto-oncogene and induce apoptosis. Oncogene 22, 2772–2781 (2003). https://doi.org/10.1038/sj.onc.1206359

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