LARP7-Mediated U6 snRNA Modification Ensures Splicing Fidelity and Spermatogenesis in Mice.

Mol. Cell. 2020 Mar 05;77(5):999-1013.e6. Epub 2020 Feb 03

Xin Wang ¹ , Zhi-Tong Li ¹ , Yue Yan ¹ , Penghui Lin ² , Wei Tang ³ ,

Xin Wang ¹ , Zhi-Tong Li ¹ , Yue Yan ¹ , Penghui Lin ² , Wei Tang ³ , Daniele Hasler ⁴ , Rajyalakshmi Meduri ⁵ , Ye Li ⁶ , Min-Min Hua ⁷ , Hui-Tao Qi ¹ , Di-Hang Lin ¹ , Hui-Juan Shi ⁸ , Jingyi Hui ¹ , Jinsong Li ⁹ , Dangsheng Li ¹ , Jian-Hua Yang ² , Jinzhong Lin ⁶ , Gunter Meister ⁴ , Utz Fischer ⁵ , Mo-Fang Liu ¹⁰

+ et al

Author information

¹ State Key Laboratory of Molecular Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences - University of Chinese Academy of Sciences, Shanghai 200031, China.
² Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, Guangdong 510275, China.
³ Animal Core Facility, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
⁴ Laboratory for RNA Biology, Biochemistry Center Regensburg (BZR), University of Regensburg, Regensburg, Germany.
⁵ Department of Biochemistry, University of Würzburg, 97074 Würzburg, Germany.
⁶ State Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200438, China.
⁷ State Key Laboratory of Molecular Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences - University of Chinese Academy of Sciences, Shanghai 200031, China; NHC Key Lab of Reproduction Regulation (Shanghai Institute of Planned Parenthood Research), Pharmacy School, Fudan University, Shanghai 200032, China.
⁸ NHC Key Lab of Reproduction Regulation (Shanghai Institute of Planned Parenthood Research), Pharmacy School, Fudan University, Shanghai 200032, China.
⁹ State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China; School of Life Science and Technology, Shanghai Tech University, Shanghai 201210, China.
¹⁰ State Key Laboratory of Molecular Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences - University of Chinese Academy of Sciences, Shanghai 200031, China; School of Life Science and Technology, Shanghai Tech University, Shanghai 201210, China. Electronic address: mfliu@sibcb.ac.cn.

摘要

U6 snRNA, as an essential component of the catalytic core of the pre-mRNA processing spliceosome, is heavily modified post-transcriptionally, with 2'-O-methylation being most common. The role of these modifications in pre-mRNA splicing as well as their physiological function in mammals have remained largely unclear. Here we report that the La-related protein functions as a critical cofactor for 2'-O-methylation of U6 in mouse male germ cells. Mechanistically, Lduanyu377 promotes U6 loading onto box C/D snoRNP, facilitating U6 2'-O-methylation by box C/D snoRNP. Importantly, ablation of Lduanyu377 in the male germline causes defective U6 2'-O-methylation, massive alterations in pre-mRNA splicing, and spermatogenic failure in mice, which can be rescued by ectopic expression of wild-type Lduanyu377 but not an U6-loading-deficient mutant Our data uncover a novel role of Lduanyu377 in regulating U6 2'-O-methylation and demonstrate the functional requirement of such modification for splicing fidelity and spermatogenesis in mice.

KEYWORDS: 2′-O-methylation, LARP7, U6 snRNA, box C/D snoRNP, pre-mRNA splicing, spermatogenesis

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