Department of Developmental Biochemistry, University of Goettingen, 37077 Goettingen, Germany.
Department for Molecular Structural Biology, University of Goettingen, 37077 Goettingen, Germany.
Biomolecules. 2021 Oct 13;11(10):1507. doi: 10.3390/biom11101507.
Many multicellular organisms specify germ cells during early embryogenesis by the inheritance of ribonucleoprotein (RNP) granules known as germplasm. However, the role of complex interactions of RNP granules during germ cell specification remains elusive. This study characterizes the interaction of RNP granules, Buc, and zebrafish Vasa (zfVasa) during germ cell specification. We identify a novel zfVasa-binding motif (Buc-VBM) in Buc and a Buc-binding motif (zfVasa-BBM) in zfVasa. Moreover, we show that Buc and zfVasa directly bind in vitro and that this interaction is independent of the RNA. Our circular dichroism spectroscopy data reveal that the intrinsically disordered Buc-VBM peptide forms alpha-helices in the presence of the solvent trifluoroethanol. Intriguingly, we further demonstrate that Buc-VBM enhances zfVasa ATPase activity, thereby annotating the first biochemical function of Buc as a zfVasa ATPase activator. Collectively, these results propose a model in which the activity of zfVasa is a central regulator of primordial germ cell (PGC) formation and is tightly controlled by the germplasm organizer Buc.
许多多细胞生物通过核糖核蛋白（RNP）颗粒的遗传，即生殖质，在胚胎早期指定生殖细胞。然而，RNP 颗粒的复杂相互作用在生殖细胞指定中的作用仍然难以捉摸。本研究描述了 RNP 颗粒、Buc 和斑马鱼 Vasa（zfVasa）在生殖细胞指定过程中的相互作用。我们在 Buc 中鉴定出一个新的 zfVasa 结合基序（Buc-VBM）和 zfVasa 中的 Buc 结合基序（zfVasa-BBM）。此外，我们表明 Buc 和 zfVasa 可以直接在体外结合，并且这种相互作用不依赖于 RNA。我们的圆二色性光谱数据显示， intrinsically disordered Buc-VBM 肽在三氟乙醇溶剂存在下形成 alpha-螺旋。有趣的是，我们进一步证明 Buc-VBM 增强了 zfVasa 的 ATP 酶活性，从而将 Buc 作为 zfVasa ATP 酶激活剂的第一个生化功能注释。总的来说，这些结果提出了一个模型，即 zfVasa 的活性是原始生殖细胞（PGC）形成的中心调节剂，并且受到生殖质组织者 Buc 的严格控制。
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