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首页> 美国卫生研究院文献>Molecular and Cellular Biology >An abundant high-mobility-group-like protein is targeted to micronuclei in a cell cycle-dependent and developmentally regulated fashion in Tetrahymena thermophila.
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An abundant high-mobility-group-like protein is targeted to micronuclei in a cell cycle-dependent and developmentally regulated fashion in Tetrahymena thermophila.

机译：嗜热四膜膜虫以细胞周期依赖性和发育调控的方式将大量的高迁移率族样蛋白靶向微核。

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In this report, we have demonstrated for the first time that an abundant high-mobility-group (HMG)-like protein, HMG B, previously thought to be specific to macronuclei in Tetrahymena thermophila, is also present in micronuclei. Biochemical data document the fact that HMG B is extremely labile in micronuclei. Unless extreme precautions are taken during the isolation of nuclei (addition of 1% formaldehyde to the nucleus isolation buffer), HMG B is not detected in micronuclei. Using polyclonal antibodies highly selective for HMG B, immunoblotting and immunofluorescence analyses show that the presence of HMG B in micronuclei is dynamic, correlating well with known periods of micronuclear DNA replication. This is the case not only during the vegetative cell cycle but also during early stages of the sexual cycle, conjugation, when the presence of HMG B in micronuclei is also closely correlated with meiotic DNA recombination and repair. Since micronuclei are transcriptionally inactive during vegetative growth, our data lend support to the idea that HMG B does not function exclusively in the establishment of transcriptionally competent chromatin. However, micronuclei are transcriptionally active during early stages of conjugation. Evidence that HMG B is strongly synthesized and deposited into micronuclei during this stage is presented. Therefore, it is tempting to suggest that HMG B may play an important role in remodeling micronuclear chromatin into an "active," more open configuration. We favor a model wherein HMG B, like other abundant, low-specificity HMG box-containing proteins, functions to wrap DNA, presumably modulating higher-order chromatin structure for a broad range of biological processes, including transcription and replication.
机译：在本报告中，我们首次证明了微核中还存在大量高迁移率族（HMG）样蛋白HMG B，以前认为该蛋白对嗜热四膜膜虫的大核具有特异性。生化数据记录了HMG B在微核中极度不稳定的事实。除非在分离核过程中采取了极端的预防措施（向核分离缓冲液中添加1％甲醛），否则在微核中不会检测到HMGB。使用对HMG B具有高度选择性的多克隆抗体，免疫印迹和免疫荧光分析表明HMG B在微核中的存在是动态的，与微核DNA复制的已知时期密切相关。这种情况不仅发生在营养细胞周期中，而且在性周期的早期共轭中也是如此，此时微核中HMG B的存在也与减数分裂DNA重组和修复密切相关。由于微核在营养生长过程中是无转录活性的，因此我们的数据支持了HMG B并非仅在建立转录感受态染色质中起作用的观点。但是，在结合的早期阶段，微核具有转录活性。证据表明，在此阶段，HMG B被强烈合成并沉积在微核中。因此，很容易暗示HMG B可能在将微核染色质重塑为“活性的”更开放的构型中起重要作用。我们喜欢这样一种模型，其中HMG B像其他大量的，低特异性的HMG盒蛋白一样，具有包裹DNA的功能，推测是在更广泛的生物学过程（包括转录和复制）中调节高级染色质结构。

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