TY - JOUR
T1 - MIWI2 as an Effector of DNA Methylation and Gene Silencing in Embryonic Male Germ Cells
AU - Kojima-Kita, Kanako
AU - Kuramochi-Miyagawa, Satomi
AU - Nagamori, Ippei
AU - Ogonuki, Narumi
AU - Ogura, Atsuo
AU - Hasuwa, Hidetoshi
AU - Akazawa, Takashi
AU - Inoue, Norimitsu
AU - Nakano, Toru
N1 - Funding Information:
The authors thank Dr. S. Chuma of Kyoto University for the anti-TDRD9 antibody, Dr. S. Tajima of Osaka University for the anti-Dnmt3a antibody and the FLAG-tagged-Dnmt3a and Dnmt3a2 expression plasmids inserted into the pcDNA4 vector, and Ms. Y. Esaki for assistance in generating the Tg mice. We also thank Ms. N. Asada and NPO Biotechnology Research and Development for the technical assistance and Ms. M. Imaizumi for the secretarial work. This work was supported in part by a Grant-in-Aid for Scientific Research Grant on Innovative Areas, “Epigenome Dynamics and Regulation in Germ Cells” (26112511), Research (A) (15H02509), Research (B) (24370074) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT)/Japan Society for the Promotion of Science (JSPS) and Core Research for Evolutional Science and Technology (CREST) (J150701424) from the Japan Agency for Medical Research and Development (AMED).
Publisher Copyright:
© 2016 The Authors
PY - 2016/9/13
Y1 - 2016/9/13
N2 - During the development of mammalian embryonic germ cells, global demethylation and de novo DNA methylation take place. In mouse embryonic germ cells, two PIWI family proteins, MILI and MIWI2, are essential for the de novo DNA methylation of retrotransposons, presumably through PIWI-interacting RNAs (piRNAs). Although piRNA-associated MIWI2 has been reported to play critical roles in the process, its molecular mechanisms have remained unclear. To identify the mechanism, transgenic mice were produced; they contained a fusion protein of MIWI2 and a zinc finger (ZF) that recognized the promoter region of a type A LINE-1 gene. The ZF-MIWI2 fusion protein brought about DNA methylation, suppression of the type A LINE-1 gene, and a partial rescue of the impaired spermatogenesis of MILI-null mice. In addition, ZF-MIWI2 was associated with the proteins involved in DNA methylation. These data indicate that MIWI2 functions as an effector of de novo DNA methylation of the retrotransposon.
AB - During the development of mammalian embryonic germ cells, global demethylation and de novo DNA methylation take place. In mouse embryonic germ cells, two PIWI family proteins, MILI and MIWI2, are essential for the de novo DNA methylation of retrotransposons, presumably through PIWI-interacting RNAs (piRNAs). Although piRNA-associated MIWI2 has been reported to play critical roles in the process, its molecular mechanisms have remained unclear. To identify the mechanism, transgenic mice were produced; they contained a fusion protein of MIWI2 and a zinc finger (ZF) that recognized the promoter region of a type A LINE-1 gene. The ZF-MIWI2 fusion protein brought about DNA methylation, suppression of the type A LINE-1 gene, and a partial rescue of the impaired spermatogenesis of MILI-null mice. In addition, ZF-MIWI2 was associated with the proteins involved in DNA methylation. These data indicate that MIWI2 functions as an effector of de novo DNA methylation of the retrotransposon.
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U2 - 10.1016/j.celrep.2016.08.027
DO - 10.1016/j.celrep.2016.08.027
M3 - Article
C2 - 27626653
AN - SCOPUS:84991690899
SN - 2211-1247
VL - 16
SP - 2819
EP - 2828
JO - Cell Reports
JF - Cell Reports
IS - 11
ER -