Department of Molecular Biology and Biochemistry, Wesleyan University, Middletown, Connecticut, United States of America.
PLoS Genet. 2013;9(10):e1003837. doi: 10.1371/journal.pgen.1003837. Epub 2013 Oct 3.
The synaptonemal complex (SC) is a widely conserved structure that mediates the intimate alignment of homologous chromosomes during meiotic prophase and is required for proper homolog segregation at meiosis I. However, fundamental details of SC architecture and assembly remain poorly understood. The coiled-coil protein, Zip1, is the only component whose arrangement within the mature SC of budding yeast has been extensively characterized. It has been proposed that the Small Ubiquitin-like MOdifier, SUMO, plays a role in SC assembly by linking chromosome axes with Zip1's C termini. The role of SUMO in SC structure has not been directly tested, however, because cells lacking SUMO are inviable. Here, we provide direct evidence for SUMO's function in SC assembly. A meiotic smt3 reduction-of-function strain displays reduced sporulation, abnormal levels of crossover recombination, and diminished SC assembly. SC structures are nearly absent when induced at later meiotic time points in the smt3 reduction-of-function background. Using Structured Illumination Microscopy we furthermore determine the position of SUMO within budding yeast SC structure. In contrast to previous models that positioned SUMO near Zip1's C termini, we demonstrate that SUMO lies at the midline of SC central region proximal to Zip1's N termini, within a subdomain called the "central element". The recently identified SUMOylated SC component, Ecm11, also localizes to the SC central element. Finally, we show that SUMO, Ecm11, and even unSUMOylatable Ecm11 exhibit Zip1-like ongoing incorporation into previously established SCs during meiotic prophase and that the relative abundance of SUMO and Ecm11 correlates with Zip1's abundance within SCs of varying Zip1 content. We discuss a model in which central element proteins are core building blocks that stabilize the architecture of SC near Zip1's N termini, and where SUMOylation may occur subsequent to the incorporation of components like Ecm11 into an SC precursor structure.
联会复合体(SC)是一种广泛保守的结构,它介导同源染色体在减数分裂前期的紧密排列,并在减数分裂 I 中正确分离同源染色体。然而,SC 结构和组装的基本细节仍然知之甚少。螺旋蛋白 Zip1 是唯一一种在出芽酵母成熟 SC 中排列方式得到广泛描述的成分。有人提出,小泛素样修饰物 SUMO 通过将染色体轴与 Zip1 的 C 末端连接起来,在 SC 组装中发挥作用。然而,SUMO 在 SC 结构中的作用尚未得到直接测试,因为缺乏 SUMO 的细胞是不可存活的。在这里,我们提供了 SUMO 在 SC 组装中的作用的直接证据。一个减数分裂 smt3 功能降低的菌株显示出孢子形成减少、交叉重组异常水平和 SC 组装减少。在 smt3 功能降低的背景下,在后期减数分裂时间点诱导时,SC 结构几乎不存在。此外,我们使用结构光照明显微镜确定了 SUMO 在出芽酵母 SC 结构中的位置。与之前将 SUMO 定位在 Zip1 的 C 末端附近的模型相反,我们证明 SUMO 位于 SC 中央区域的中线附近,靠近 Zip1 的 N 末端,在一个称为“中央元件”的亚域内。最近鉴定的 SUMOylated SC 成分 Ecm11 也定位于 SC 中央元件。最后,我们表明 SUMO、Ecm11,甚至是不可 SUMOylated 的 Ecm11,都在减数分裂前期持续整合到先前建立的 SC 中,并且 SUMO 和 Ecm11 的相对丰度与 SC 中 Zip1 的丰度相关,而 SC 中 Zip1 的含量不同。我们讨论了一种模型,其中中央元件蛋白是核心构建块,它们在 Zip1 的 N 末端附近稳定 SC 的结构,并且 SUMOylation 可能发生在 Ecm11 等成分整合到 SC 前体结构之后。