Ribonuclease activity of MARF1 controls oocyte RNA homeostasis and genome integrity in mice.
Ribonuclease activity of MARF1 controls oocyte RNA homeostasis and genome integrity in mice.
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MARF1 的核糖核酸酶活性控制小鼠卵母细胞 RNA 稳态和基因组完整性
DOI：
10.1073/pnas.1809744115
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发表时间：
2018-10-30
影响因子：
11.1
通讯作者：
Su YQ
中科院分区：
文献类型：
--
作者：
Yao Q;Cao G;Li M;Wu B;Zhang X;Zhang T;Guo J;Yin H;Shi L;Chen J;Yu X;Zheng L;Ma J;Su YQ
虽然MARF1（减数分裂调节因子和mRNA稳定因子1）是一种古老的蛋白质，但其在哺乳动物雌性生殖细胞发育和生育中的功能是最近才被确定的。它对卵母细胞减数分裂的进展和抵御逆转录转座子的破坏至关重要，逆转录转座子会导致卵母细胞基因组的损伤。这些过程依赖于MARF1在卵子发生过程中单独结合RNA和作为核糖核酸酶的能力。在这里，我们揭示了使MARF1活性的分子结构和功能机制，并提供了对形成卵母细胞转录组的复杂转录后过程的见解。产生正常的卵子用于受精和物种繁殖需要完成减数分裂和保护基因组免受反转录转座子的破坏。Marf1（减数分裂调节因子和mRNA稳定因子1）的突变导致小鼠卵母细胞中这两个关键过程的缺陷，从而导致不育。预测MARF1具有核糖核酸酶活性，但MARF1功能的结构基础及其假定的核糖核酸酶结构域对突变卵母细胞表型的贡献尚不清楚。因此，我们解析了MARF1关键结构域的晶体结构，并通过生化和诱变分析证明MARF1的核糖核酸酶活性控制着卵母细胞减数分裂的进程和反转录转座子的监视。MARF1的n端NYN结构域类似于Vpa0982、T4 RNase H和MCPIP1的核酸酶结构域，包含4个保守的天冬氨酸残基D178、D215、D246和D272。MARF1的c端LOTUS结构域采用翼状螺旋-转-螺旋折叠，结合ssRNA和dsRNA。纯化的MARF1在体外切割ssrna，但这种切割活性被保守的天冬氨酸在其NYN结构域的突变和LOTUS结构域的截断所消除。此外，体内D272残基的点突变导致小鼠出现雌性不育表型，导致卵母细胞减数分裂恢复失败，Line1和Iap反转录子转录物升高，DNA双链断裂。因此，MARF1核糖核酸酶活性控制卵母细胞减数分裂和基因组完整性。这种活性取决于催化NYN结构域中保守的天冬氨酸残基和LOTUS结构域的rna结合活性。
Significance Although MARF1 (meiosis regulator and mRNA stability factor 1) is an ancient protein, identification of its function in mammalian female germ cell development and fertility is recent. It is crucial for the progression of oocyte meiosis and defense against the ravages of retrotransposons, which can cause damage to the oocyte’s genome. These processes are dependent upon the ability of MARF1 to act alone both to bind RNA and to function as a ribonuclease during oogenesis. Here we reveal the molecular structure and functional mechanisms that enable MARF1 activity and provide insight into the complex posttranscriptional processes that shape the oocyte transcriptome. Producing normal eggs for fertilization and species propagation requires completion of meiosis and protection of the genome from the ravages of retrotransposons. Mutation of Marf1 (meiosis regulator and mRNA stability factor 1) results in defects in both these key processes in mouse oocytes and thus in infertility. MARF1 was predicted to have ribonuclease activity, but the structural basis for the function of MARF1 and the contribution of its putative ribonuclease domain to the mutant oocyte phenotype was unknown. Therefore, we resolved the crystal structures of key domains of MARF1 and demonstrated by biochemical and mutagenic analyses that the ribonuclease activity of MARF1 controls oocyte meiotic progression and retrotransposon surveillance. The N-terminal NYN domain of MARF1 resembles the nuclease domains of Vpa0982, T4 RNase H, and MCPIP1 and contains four conserved aspartate residues, D178, D215, D246, and D272. The C-terminal LOTUS domain of MARF1 adopts a winged helix-turn-helix fold and binds ssRNA and dsRNA. Purified MARF1 cleaved ssRNAs in vitro, but this cleavage activity was abolished by mutations of conserved aspartates in its NYN domain and truncation of the LOTUS domain. Furthermore, a point mutation in the D272 residue in vivo caused a female-only infertile phenotype in mice, with failure of meiotic resumption and elevation of Line1 and Iap retrotransposon transcripts and DNA double-strand breaks in oocytes. Therefore, the ribonuclease activity of MARF1 controls oocyte meiosis and genome integrity. This activity depends upon conserved aspartic residues in the catalytic NYN domain and the RNA-binding activity of the LOTUS domain.
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影响因子：
11.4
作者：
Lau, Nelson C.;Ohsumi, Toshiro;Blower, Michael D.
通讯作者：
Blower, Michael D.
DOI：
10.1073/pnas.0911640106
发表时间：
2009-12-01
影响因子：
11.1
作者：
Chen, Chen;Jin, Jing;Pawson, Tony
通讯作者：
Pawson, Tony
果蝇种系诱导剂 Oskar 的晶体结构确定了具有不同 Vasa 螺旋酶和 RNA 结合活性的两个结构域
影响因子：
8.8
作者：
Jeske, Mandy;Bordi, Matteo;Ephrussi, Anne
通讯作者：
Ephrussi, Anne
DOI：
10.1261/rna.060939.117
发表时间：
2017-11
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Gainetdinov I;Skvortsova Y;Kondratieva S;Funikov S;Azhikina T
通讯作者：
Azhikina T
影响因子：
64.8
作者：
Morgan M;Much C;DiGiacomo M;Azzi C;Ivanova I;Vitsios DM;Pistolic J;Collier P;Moreira PN;Benes V;Enright AJ;O'Carroll D
通讯作者：
O'Carroll D