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窦卵泡来源的人卵母细胞中RNA聚合酶II的核分布:相对于转录状态的动力学以及与剪接因子的关联

Nuclear distribution of RNA polymerase II in human oocytes from antral follicles: dynamics relative to the transcriptional state and association with splicing factors.

作者信息

Parfenov V N, Davis D S, Pochukalina G N, Kostyuchek D, Murti K G

机构信息

Institute of Cytology, Russian Academy of Science, St. Petersburg, Russia.

出版信息

J Cell Biochem. 2000 Apr;77(4):654-65.

Abstract

The intranuclear distribution of two (unphosphorylated and hyperphosphorylated) forms of RNA polymerase II (Pol II) was studied in human oocytes from antral follicles using immunogold labeling/electron microscopy. The distribution of Pol II was analyzed relative to the transcriptional state of the oocyte as well as to the distribution of two splicing factors (snRNPs and SC-35) in the intranuclear entities, namely, interchromatin granule clusters (IGCs), nucleolus-like bodies (NLBs), and perichromatin fibrils (PFs). The results showed that (1) antibodies directed against two forms of Pol II have similar pattern of intranuclear distribution, (2) both Pol II and splicing factors progressively accumulate in IGCs with decrease in the transcriptional activity of the oocyte nucleus, (3) both Pol II and splicing factors localize to PFs, and (4) Pol II is present in the NLBs at all transcriptional states of the oocyte nucleus. These studies confirm earlier proposals that PFs represent a nuclear domain in which RNA transcription/processing are spatially coupled. The accumulation of Pol II and splicing factors in IGCs concomitant with a decrease in the transcriptional activity suggests a coordinated mechanism for the movement of both Pol II and splicing factors from the sites of action to the sites of storage.

摘要

利用免疫金标记/电子显微镜技术,研究了来自窦状卵泡的人类卵母细胞中两种形式(未磷酸化和高度磷酸化)的RNA聚合酶II(Pol II)的核内分布。分析了Pol II相对于卵母细胞转录状态以及两种剪接因子(小核核糖核蛋白和SC-35)在核内实体(即染色质间颗粒簇(IGC)、核仁样体(NLB)和染色质周纤维(PF))中的分布情况。结果表明:(1)针对两种形式Pol II的抗体具有相似的核内分布模式;(2)随着卵母细胞核转录活性降低,Pol II和剪接因子均在IGC中逐渐积累;(3)Pol II和剪接因子均定位于PF;(4)在卵母细胞核的所有转录状态下,Pol II均存在于NLB中。这些研究证实了早期的观点,即PF代表一个RNA转录/加工在空间上偶联的核结构域。Pol II和剪接因子在IGC中的积累与转录活性降低相伴,提示了Pol II和剪接因子从作用位点向储存位点移动的协调机制。

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