Anderson L K, Offenberg H H, Verkuijlen W M, Heyting C
Department of Genetics, Wageningen Agricultural University, Dreijenlaan 2, 6703 HA Wageningen, The Netherlands.
Proc Natl Acad Sci U S A. 1997 Jun 24;94(13):6868-73. doi: 10.1073/pnas.94.13.6868.
Early meiotic nodules (also called recombination nodules) are proteinaceous structures about 100 nm in diameter that are associated with forming synaptonemal complexes (SCs) during early prophase I of meiosis. Although their function is unknown, early nodules may be involved in searching for DNA homology before synaptic initiation. Two potential components of early nodules are Rad51 and Dmc1 proteins. These proteins are important for meiotic recombination in eukaryotes and are homologous to RecA, the major protein that catalyzes homologous pairing and DNA strand exchange in prokaryotes. In addition, Rad51 has been localized by immunofluorescence in abundant foci that may correspond to early nodules in yeast, lily, and mouse. In yeast and lily, Dmc1 and Lim15, the lily homolog of Dmc1, colocalize with Rad51. Here, using electron microscopic immunogold localization to spreads of zygotene and early pachytene SCs from lily, we confirm that RecA-like proteins are components of early nodules. The antibody used was generated to full-length tomato Rad51 protein and binds to both Rad51 and Lim15 in immunoblots of lily primary microsporocyte proteins. The labeled early nodules are heterogeneous in size and are associated with both axial elements and SCs. There are two classes of early nodules, those that are densely labeled with gold and those that are not labeled at all. This result may be due to technical limitations associated with using spread preparations or to differences in the nodules themselves. The presence of Rad51 and/or Lim15 proteins in early nodules supports the hypothesis that early nodules are involved in recombination-related events during meiosis.
早期减数分裂结节(也称为重组结节)是直径约100纳米的蛋白质结构,在减数分裂前期I早期与联会复合体(SC)的形成相关。尽管其功能尚不清楚,但早期结节可能在突触起始前参与寻找DNA同源性。早期结节的两个潜在成分是Rad51和Dmc1蛋白。这些蛋白质对真核生物的减数分裂重组很重要,并且与RecA同源,RecA是催化原核生物中同源配对和DNA链交换的主要蛋白质。此外,通过免疫荧光法已将Rad51定位在丰富的病灶中,这些病灶可能对应于酵母、百合和小鼠中的早期结节。在酵母和百合中,Dmc1和Dmc1的百合同源物Lim15与Rad51共定位。在这里,我们使用电子显微镜免疫金定位技术对百合偶线期和早粗线期SC的铺展进行研究,证实了类RecA蛋白是早期结节的成分。所用抗体是针对全长番茄Rad51蛋白产生的,在百合初级小孢子母细胞蛋白的免疫印迹中与Rad51和Lim15都结合。标记的早期结节大小不一,与轴元件和SC都有关联。早期结节有两类,一类被金标记得很密集,另一类则完全没有被标记。这个结果可能是由于使用铺展制剂相关的技术限制,或者是结节本身的差异。早期结节中存在Rad51和/或Lim15蛋白支持了早期结节参与减数分裂期间与重组相关事件的假说。