Random mutagenesis of PDZ(Omi) domain and selection of mutants that specifically bind the Myc proto-oncogene and induce apoptosis

    Authors

    D. Junqueira; L. Cilenti; L. Musumeci; J. M. Sedivy;A. S. Zervos

    Comments

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    Abbreviated Journal Title

    Oncogene

    Keywords

    Omi/HtrA2; PDZ domains; protein-protein interactions; protein design; Myc proto-oncogene; apoptosis; yeast two-hybrid system; TUMOR-SUPPRESSOR PROTEIN; SERINE-PROTEASE; PDZ DOMAINS; C-MYC; CELL-DEATH; MOLECULAR CHARACTERIZATION; TISSUE DISTRIBUTION; SEPTATE; JUNCTIONS; CRYSTAL-STRUCTURE; MEMBRANE PROTEIN; Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics &; Heredity

    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 beta-finger are also reported to bind some PDZ domains. Using a modified yeast two-hybrid system, PDZ(Omi) 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 PDZ(m) 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.

    Journal Title

    Oncogene

    Volume

    22

    Issue/Number

    18

    Publication Date

    1-1-2003

    Document Type

    Article

    Language

    English

    First Page

    2772

    Last Page

    2781

    WOS Identifier

    WOS:000182569300008

    ISSN

    0950-9232

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