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      Associations between PIWI proteins and TDRD1/MTR-1 are critical for integrated subcellular localization in murine male germ cells.

      Genes to Cells
      Animals, Argonaute Proteins, Blotting, Western, Cell Line, Cell Nucleus, metabolism, Cytoplasm, Green Fluorescent Proteins, genetics, Humans, Immunohistochemistry, Immunoprecipitation, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Microscopy, Electron, Mutation, Protein Binding, Proteins, Recombinant Fusion Proteins, Ribonucleoproteins, Small Nuclear, Spermatocytes, cytology, ultrastructure, Testis, Transfection

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          Abstract

          The mouse Piwi family proteins (MILI, MIWI and MIWI2) play pivotal roles in spermatogenesis through transcriptional and post-transcriptional gene regulation. To reveal the molecular functions of these proteins, we investigate the proteins that bind to MILI in adult mouse testes. We found that both MILI and MIWI bind to TDRD1/MTR-1, which is also an essential protein for spermatogenesis. Co-immunoprecipitation assays and subcellular localization of the proteins and mutants thereof revealed a complex formation involving MILI, MIWI and TDRD1/MTR-1. In addition, the subcellular localizations of MILI and TDRD1/MTR-1 were altered, and chromatoid body formation was impaired in the MIWI-null round spermatids. These data suggest that the formation of complexes between MILI, MIWI and TDRD1/MTR-1 is critical for the integrated subcellular localizations of these proteins, and is presumably essential for spermatogenesis.

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