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Cytological analysis of MRE11 protein during early meiotic prophase I in Arabidopsis and tomato

机译:拟南芥和番茄减数分裂前期I期MRE11蛋白的细胞学分析

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摘要

Early recombination nodules (ENs) are multiprotein complexes that are thought to be involved in synapsis and recombination, but little is known about their components or how they may be involved in these events. In this study, we describe the cytological behavior of a possible EN component, MRE11, a protein that is important for the repair of the numerous, programmed deoxyribonucleic acid double-strand breaks (DSBs) that occur early in the meiotic prophase. By immunofluorescence, many MRE11 foci were associated with chromosomal axes during early prophase I in both wild-type Arabidopsis and tomato primary microsporocytes. Similar patterns of MRE11 foci were observed in two Arabidopsis mutants (Atspo11-1 and Atprd1) that are defective in DSB formation and synapsis. In tomato chromosomes, MRE11 foci were more common in distal euchromatin than in proximal heterochromatin, consistent with known EN patterns. However, electron microscopic immunogold localization demonstrated that only about 10% of ENs were labeled, and most MRE11 label was associated with synaptonemal complex components. Thus, in plants, MRE11 foci are not dependent on DSB formation, and most MRE11 foci do not correspond to ENs. More generally, our results show that the simple presence of large numbers of fluorescent foci associated with synapsing chromosomes is insufficient evidence to equate these foci with ENs.
机译:早期重组结节(ENs)是一种多蛋白复合物,被认为与突触和重组有关,但对其成分或如何参与这些事件知之甚少。在这项研究中,我们描述了可能的EN成分MRE11的细胞学行为,该蛋白质对于减数分裂前期早期发生的许多程序化脱氧核糖核酸双链断裂(DSB)的修复非常重要。通过免疫荧光,在野生型拟南芥和番茄原代微孢子细胞中,许多MRE11病灶在前期I早期都与染色体轴相关。在两个拟南芥突变体(Atspo11-1和Atprd1)中观察到了相似的MRE11基因座模式,这些突变体在DSB的形成和突触中存在缺陷。在番茄染色体中,远端常染色质中的MRE11病灶比近端异染色质中的更常见,这与已知的EN模式一致。但是,电子显微镜免疫金定位表明,只有约10%的EN被标记,大多数MRE11标记与突触复合物相关。因此,在植物中,MRE11焦点不依赖于DSB的形成,大多数MRE11焦点不对应于EN。更普遍地,我们的结果表明,与突触染色体相关的大量荧光灶的简单存在不足以证明这些灶与EN等同。

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