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¹ Department of Pathology, Medical School, Osaka University, Yamada-oka 2-2 Suita, Osaka 565-0871, Japan
² PREST, JST, 4-1-8 Honcho Kawaguchi, Saitama, Japan
³ Institute for Frontier Medical Sciences, and Graduate School of Medicine, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan

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.

^* Correspondence: tnakano{at}patho.med.osaka-u.ac.jp or smiya{at}patho.med.osaka-u.ac.jp