Discipline of Biochemistry, School of Molecular and Biomedical Science, University of Adelaide, Adelaide, Australia.
PLoS One. 2011;6(6):e20773. doi: 10.1371/journal.pone.0020773. Epub 2011 Jun 13.
The ability of germ cells to carry out a gene regulatory program distinct from the surrounding somatic tissue, and their capacity to specify an entire new organism has made them a focus of many studies that seek to understand how specific regulatory mechanisms, particularly post-transcriptional mechanisms, contribute to cell fate. In zebrafish, germ cells are specified through the inheritance of cytoplasmic determinants, termed the germ plasm, which contains a number of maternal mRNAs and proteins. Investigation of several of these messages has revealed that the restricted localisation of these mRNAs to the germ plasm and subsequent germ cells is due to cis-acting sequence elements present in their 3'UTRs. Here we show that a member of the Hu family of RNA-binding proteins, HuB, is maternally provided in the zebrafish embryo and exhibits germ cell specific expression during embryogenesis. Restriction of HuB mRNA to the germ cells is dependent on a number of sequence elements in its 3'UTR, which act to degrade the mRNA in the soma and stabilise it in the germ cells. In addition, we show that the germ cell specific RNA-binding protein DAZL is able to promote HuB mRNA stability and translation in germ cells, and further demonstrate that these activities require a 30 nucleotide element in the 3'UTR. Our study suggests that DAZL specifically binds the HuB 3'UTR and protects the message from degradation and/or enhances HuB translation, leading to the germ cell specific expression of HuB protein.
生殖细胞能够执行与周围体细胞组织不同的基因调控程序,并且能够指定整个新生物体,这使它们成为许多研究的焦点,这些研究旨在了解特定的调控机制,特别是转录后机制,如何促进细胞命运。在斑马鱼中,生殖细胞是通过细胞质决定因素的遗传来指定的,这些细胞质决定因素被称为生殖质,其中包含许多母体 mRNA 和蛋白质。对其中一些信息的研究表明,这些 mRNA 局限于生殖质和随后的生殖细胞中的特定局部化是由于其 3'UTR 中存在顺式作用序列元件。在这里,我们表明 Hu 家族的 RNA 结合蛋白之一 HuB 在斑马鱼胚胎中是母体提供的,并在胚胎发生过程中表现出生殖细胞特异性表达。HuB mRNA 在生殖细胞中的限制依赖于其 3'UTR 中的许多序列元件,这些元件在体细胞中降解 mRNA 并稳定其在生殖细胞中。此外,我们表明,生殖细胞特异性 RNA 结合蛋白 DAZL 能够促进 HuB mRNA 在生殖细胞中的稳定性和翻译,并且进一步证明这些活性需要 3'UTR 中的 30 个核苷酸元件。我们的研究表明,DAZL 特异性结合 HuB 3'UTR 并保护该消息免受降解和/或增强 HuB 翻译,导致 HuB 蛋白在生殖细胞中的特异性表达。