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YTHDC2 regulates spermatogenesis through promoting the translation of N 6 ‐methyladenosine‐modified RNA
Author(s) -
Hsu Phillip J,
Zhu Yunfei,
Ma Honghui,
Cui Yiqiang,
Shi Xiaodan,
Luo Guanzheng,
Lu Zhike,
Shi Hailing,
Dai Qing,
Clark Marcus,
Shen Bin,
He Chuan
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.595.10
Subject(s) - messenger rna , rna binding protein , rna splicing , rna helicase a , microbiology and biotechnology , translation (biology) , spermatogenesis , biology , rna , genetics , gene , helicase , endocrinology
N 6 ‐methyladenosine (m 6 A) is the most common internal modification in eukaryotic mRNA. It is dynamically installed and removed, and acts as an essential layer of mRNA metabolism, regulating biological processes including stem cell pluripotency, cell differentiation, and energy homeostasis. m 6 A is recognized by selective binding proteins; YTHDF1 promotes the translation of m 6 A‐containing mRNAs, YTHDF2 expedites mRNA decay, and YTHDC1 affects the splicing of its targets. The biological function of YTHDC2, another member of the YTH protein family, remains unknown. We report that YTHDC2 selectively binds m 6 A along its consensus motif GGACU. YTHDC2 promotes translation of its targets, and associates with cellular fractions involved in translation initiation. YTHDC2 contains highly conserved helicase domains, whose activity is required for its function in promoting translation. Ythdc2 knockout mice are infertile and have significantly smaller testes compared to those of littermates. In the testes, Ythdc2 is temporally expressed as meiosis begins, and germ cells of Ythdc2 knockout mice do not develop past the spermatogonium stage. Thus, YTHDC2 is an m 6 A binding protein that plays essential roles in spermatogenesis. Support or Funding Information The work was supported by the National Institutes of Health grants GM71440 and GM113194 to C.H. C.H. is an investigator of the Howard Hughes Medical Institute (HHMI). P.J.H. is supported by the University of Chicago Medical Scientist Training Program under M.C. H.M. is supported by the Postdoctoral International Exchange Program of the China Postdoctoral Council (CPC). The Mass Spectrometry Facility of the University of Chicago is funded by the National Science Foundation (CHE‐1048528). 1YTHDC2 is an m 6 A reader. A) Gel shift assay measuring the dissociation (K − d , nM, indicated at the upper left corner of the gel) of FLAG‐tagged YTHDC2 with methylated and unmethylated RNA probes. Protein concentrations are listed in nM. 4 nmol RNA probe was used. B) In vitro probe pulldown assay comparing amount of YTHDC2 protein pulled down by biotin‐labeled methylated and unmethylated RNA probes, with or without a GG (A or m 6 A) CU motif.2YTHDC2 is essential for spermatogenesis. A) Testes of Ythdc2−/− and control mice. B) Staining of testes cross sections of Ythdc2−/− and control mice. Germ cells of Ythdc2 knockout mice do not develop past the spermatogonium stage.