Leyden Melissa R, Michalik Peter, Baruffaldi Luciana, Mahmood Susheen, Kalani Ladan, Hunt Donald F, Eirin-Lopez Jose Maria, Andrade Maydianne C B, Shabanowitz Jeffrey, Ausió Juan
Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, USA.
Zoologisches Institut und Museum, Universität Greifswald, Greifswald, Germany.
bioRxiv. 2024 Jun 5:2024.06.04.597381. doi: 10.1101/2024.06.04.597381.
While there is extensive information about sperm nuclear basic proteins (SNBP) in vertebrates, there is very little information about Arthropoda by comparison. This paper aims to contribute to filling this gap by analyzing these proteins in the sperm of the noble false widow spider (Order Araneae, Family Theridiidae). To this end, we have developed a protein extraction method that allows the extraction of cysteine-containing protamines suitable for the preparation and analysis of SNBPs from samples where the amount of starting tissue material is limited. We carried out top-down mass spectrometry sequencing and molecular phylogenetic analyses to characterize the protamines of and other spiders. We also used electron microscopy to analyze the chromatin organization of the sperm, and we found it to exhibit liquid-liquid phase spinodal decomposition during the late stages of spermiogenesis. These studies further our knowledge of the distribution of SNBPs within the animal kingdom and provide additional support for a proposed evolutionary origin of many protamines from a histone H1 (H5) replication-independent precursor.
虽然关于脊椎动物精子核碱性蛋白（SNBP）有大量信息，但相比之下，关于节肢动物的信息却非常少。本文旨在通过分析高贵的寡妇蜘蛛（蜘蛛目，球蛛科）精子中的这些蛋白质来填补这一空白。为此，我们开发了一种蛋白质提取方法，该方法能够从起始组织材料量有限的样本中提取适合制备和分析SNBP的含半胱氨酸的鱼精蛋白。我们进行了自上而下的质谱测序和分子系统发育分析，以表征高贵的寡妇蜘蛛和其他蜘蛛的鱼精蛋白。我们还使用电子显微镜分析了精子的染色质组织，发现其在精子发生后期表现出液-液相旋节线分解。这些研究进一步加深了我们对SNBP在动物界分布的了解，并为许多鱼精蛋白起源于与组蛋白H1（H5）复制无关的前体这一进化起源假说提供了更多支持。
