Advanced ICT Research Institute Kobe, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe, 651-2492, Japan.
Graduate School of Frontier Biosciences, Osaka University, Suita, 565-0871, Japan.
Chromosoma. 2021 Sep;130(2-3):149-162. doi: 10.1007/s00412-021-00757-w. Epub 2021 Apr 7.
The structure of chromosomes dramatically changes upon entering meiosis to ensure the successful progression of meiosis-specific events. During this process, a multilayer proteinaceous structure called a synaptonemal complex (SC) is formed in many eukaryotes. However, in the fission yeast Schizosaccharomyces pombe, linear elements (LinEs), which are structures related to axial elements of the SC, form on the meiotic cohesin-based chromosome axis. The structure of LinEs has been observed using silver-stained electron micrographs or in immunofluorescence-stained spread nuclei. However, the fine structure of LinEs and their dynamics in intact living cells remain to be elucidated. In this study, we performed live cell imaging with wide-field fluorescence microscopy as well as 3D structured illumination microscopy (3D-SIM) of the core components of LinEs (Rec10, Rec25, Rec27, Mug20) and a linE-binding protein Hop1. We found that LinEs form along the chromosome axis and elongate during meiotic prophase. 3D-SIM microscopy revealed that Rec10 localized to meiotic chromosomes in the absence of other LinE proteins, but shaped into LinEs only in the presence of all three other components, the Rec25, Rec27, and Mug20. Elongation of LinEs was impaired in double-strand break-defective rec12 cells. The structure of LinEs persisted after treatment with 1,6-hexanediol and showed slow fluorescence recovery from photobleaching. These results indicate that LinEs are stable structures resembling axial elements of the SC.
染色体结构在进入减数分裂时会发生剧烈变化,以确保减数分裂特异性事件的成功进行。在此过程中,许多真核生物中会形成一种称为联会复合体(SC)的多层蛋白结构。然而,在裂殖酵母 Schizosaccharomyces pombe 中,线性元件(LinEs),即与 SC 的轴向元件相关的结构,在线性轴上形成。LinEs 的结构已经通过银染电子显微镜或免疫荧光染色的铺展核观察到。然而,LinEs 的精细结构及其在完整活细胞中的动力学仍有待阐明。在这项研究中,我们使用宽场荧光显微镜进行活细胞成像,并使用 3D 结构照明显微镜(3D-SIM)对 LinEs 的核心成分(Rec10、Rec25、Rec27、Mug20)和一个 LinE 结合蛋白 Hop1 进行了研究。我们发现 LinEs 沿着染色体轴形成,并在减数分裂前期伸长。3D-SIM 显微镜显示,Rec10 在没有其他 LinE 蛋白的情况下定位于减数分裂染色体上,但只有在存在其他三个成分(Rec25、Rec27 和 Mug20)的情况下才形成 LinEs。在双链断裂缺陷 rec12 细胞中,LinEs 的伸长受到损害。LinEs 的结构在 1,6-己二醇处理后仍然存在,并表现出缓慢的光漂白荧光恢复。这些结果表明 LinEs 是类似于 SC 轴向元件的稳定结构。