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piRNA (PIWI-interacting RNA) is a germ cell-specific small RNA in which biogenesis PIWI (P-element wimpy testis) family proteins play crucial roles. MILI (mouse Piwi-like), one of the three mouse PIWI family members, is indispensable for piRNA production, DNA methylation of retrotransposons presumably through the piRNA, and spermatogenesis. The biogenesis of piRNA has been divided into primary and secondary processing pathways; in both of these MILI is involved in mice. To analyze the molecular function of MILI in piRNA biogenesis, we utilized germline stem (GS) cells, which are derived from testicular stem cells and possess a spermatogonial phenotype. We established MILI-null GS cell lines and their revertant, MILI-rescued GS cells, by introducing the Mili gene with Sendai virus vector. Comparison of wild-type, MILI-null, and MILI-rescued GS cells revealed that GS cells were quite useful for analyzing the molecular mechanisms of piRNA production, especially the primary processing pathway. We found that glycerol-3-phosphate acyltransferase 2 (GPAT2), a mitochondrial outer membrane protein for lysophosphatidic acid, bound to MILI using the cells and that gene knockdown of GPAT2 brought about impaired piRNA production in GS cells. GPAT2 is not only one of the MILI bound proteins but also a protein essential for primary piRNA biogenesis.

Citation

Yusuke Shiromoto, Satomi Kuramochi-Miyagawa, Akito Daiba, Shinichiro Chuma, Ami Katanaya, Akiko Katsumata, Ken Nishimura, Manami Ohtaka, Mahito Nakanishi, Toshinobu Nakamura, Koichi Yoshinaga, Noriko Asada, Shota Nakamura, Teruo Yasunaga, Kanako Kojima-Kita, Daisuke Itou, Tohru Kimura, Toru Nakano. GPAT2, a mitochondrial outer membrane protein, in piRNA biogenesis in germline stem cells. RNA (New York, N.Y.). 2013 Jun;19(6):803-10

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PMID: 23611983

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