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

Genes Cells. 2009 Oct;14(10):1155-65. doi: 10.1111/j.1365-2443.2009.01342.x. Epub 2009 Sep 7.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Argonaute Proteins
  • Cell Cycle Proteins
  • Piwil1 protein, mouse
  • Proteins
  • Recombinant Fusion Proteins
  • Ribonucleoproteins, Small Nuclear
  • Tdrd1 protein, mouse
  • Green Fluorescent Proteins