Oocyte-specific maternal Slbp2 is required for replication-dependent histone storage and early nuclear cleavage in zebrafish oogenesis and embryogenesis.
Oocyte-specific maternal Slbp2 is required for replication-dependent histone storage and early nuclear cleavage in zebrafish oogenesis and embryogenesis.
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DOI：
10.1261/rna.067090.118
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发表时间：
2018-12
期刊：
影响因子：
--
通讯作者：
Gui JF
中科院分区：
文献类型：
--
作者：
He WX;Wu M;Liu Z;Li Z;Wang Y;Zhou J;Yu P;Zhang XJ;Zhou L;Gui JF
在哺乳动物中，茎环结合蛋白（SLBP）是复制依赖性组蛋白mRNA代谢所必需的。斑马鱼有两个slbp1，其中slbp1是视网膜神经发生所必需的。然而，slbp2在斑马鱼中的具体表达和功能尚不清楚。在本研究中，我们首先将斑马鱼slbp2鉴定为卵母细胞特异性母源因子，然后利用CRISPR/Cas9技术生成母源合子slbp2 F3纯合突变体（MZslbp2Δ4−/−）。母体Slbp2的缺失破坏了早期的核分裂，导致MBT阶段发育停滞。slbp2转基因MZslbp2Δ4−/−胚胎可以修复发育缺陷。然而，纯合突变体MZslbp1Δ1−/−发育正常，表明slbp1在斑马鱼早期胚胎发生中是不可缺少的。通过比较WT和MZslbp2Δ4−/−胚胎之间的蛋白质组和转录组分析，我们发现了许多差异表达的蛋白质和基因。与WT胚胎相比，4种复制依赖性组蛋白H2a、H2b、H3和H4的表达均降低，而组蛋白变体h2afx在MZslbp2Δ4−/−胚胎的256细胞期和高增殖期显著增加。斑马鱼Slbp2在体外可以结合组蛋白mRNA茎环，并且通过过表达H2b可以部分修复MZslbp2Δ4−/−胚胎的缺陷。目前的数据表明，在斑马鱼卵发生过程中，母体Slbp2在复制依赖性组蛋白mrna和蛋白质的储存中起着关键作用。
Stem–loop binding protein (SLBP) is required for replication-dependent histone mRNA metabolism in mammals. Zebrafish possesses two slbps, and slbp1 is necessary for retinal neurogenesis. However, the detailed expression and function of slbp2 in zebrafish are still unknown. In this study, we first identified zebrafish slbp2 as an oocyte-specific maternal factor and then generated a maternal-zygotic slbp2 F3 homozygous mutant (MZslbp2Δ4−/−) using CRISPR/Cas9. The depletion of maternal Slbp2 disrupted early nuclear cleavage, which resulted in developmental arrest at the MBT stage. The developmental defects could be rescued in slbp2 transgenic MZslbp2Δ4−/− embryos. However, homozygous mutant MZslbp1Δ1−/− developed normally, indicating slbp1 is dispensable for zebrafish early embryogenesis. Through comparative proteome and transcriptome profiling between WT and MZslbp2Δ4−/− embryos, we identified many differentially expressed proteins and genes. In comparison with those in WT embryos, four replication-dependent histones, including H2a, H2b, H3, and H4, all reduced their expression, while histone variant h2afx significantly increased in MZslbp2Δ4−/− embryos at the 256-cell stage and high stage. Zebrafish Slbp2 can bind histone mRNA stem–loop in vitro, and the defects of MZslbp2Δ4−/− embryos can be partially rescued by overexpression of H2b. The current data indicate that maternal Slbp2 plays a pivotal role in the storage of replication-dependent histone mRNAs and proteins during zebrafish oogenesis.
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