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[关于单细胞和双细胞小鼠胚胎中卫星DNA及相关蛋白相对于核仁前体的定位]

[Localization of satellite DNA and associated protein in respect to nucleolar precursor bodies in one- and two-cell mouse embryos].

作者信息

Gavrilova E V, Kuznetsova I S, Enukashvili N I, Noniashvili E M, Dyban A P, Podgornaia O I

出版信息

Tsitologiia. 2009;51(5):455-64.

Abstract

Nucleolar precursor bodies (NPB) are characteristic structures in the nuclei of one- and two cell mouse embryos. The alignment of centromeric (CEN) and pericentromeric (periCEN) chromosome regions to the chromatin layer surrounding NPB is known. Mus musculus 4 satellite DNA (satDNA) types are known to be located in CEN region--mouse minor satellite (MiSat) and mouse satellite 3 (MS3); and periCEN region--mouse major satellite (MaSat) and mouse satellite (MS4). We determined the localization of 4 types of mouse satDNA CEN and periCEN regions and associated proteins: RNA-helicase p68, SMC3, Rad21 subunits of the cohesin complex and SYCP3 subunit of the synaptonemal complex (SC). Partially flattened nuclei of the one- and two-cell embryos and embryos treated with ocadaic acids (OA) were used. Different satDNA fragments revealed distinct domains at the surface of NPB: periCEN MaSat was always localized in NPB more internally covering almost entire surface of NPB while CEN MiSat, MS3 and periCEN MS4 showed more peripheral localization. All 4 satDNA did not cover the entire areas of the NPB, indicating the presence of other DNA sequence involved in its formation. RNA-helicase p68 and components of multiprotein cohesin and synaptonemal complexes are the necessary components of NPB. Our results support the opinion that NPB serve as a precursor of chromocenters.

摘要

核仁前体(NPB)是单细胞和双细胞小鼠胚胎细胞核中的特征性结构。已知着丝粒(CEN)和着丝粒周围(periCEN)染色体区域与围绕NPB的染色质层对齐。小家鼠4种卫星DNA(satDNA)类型已知位于CEN区域——小鼠小卫星(MiSat)和小鼠卫星3(MS3);以及着丝粒周围区域——小鼠大卫星(MaSat)和小鼠卫星(MS4)。我们确定了4种小鼠satDNA的CEN和着丝粒周围区域以及相关蛋白的定位:RNA解旋酶p68、黏连蛋白复合体的SMC3、Rad21亚基以及联会复合体(SC)的SYCP3亚基。使用了单细胞和双细胞胚胎以及经冈田酸(OA)处理的胚胎的部分扁平细胞核。不同的satDNA片段在NPB表面显示出不同的结构域:着丝粒周围的MaSat总是更内部地定位在NPB中,几乎覆盖NPB的整个表面,而CEN的MiSat、MS3和着丝粒周围的MS4显示出更外围的定位。所有4种satDNA并未覆盖NPB的整个区域,表明存在参与其形成的其他DNA序列。RNA解旋酶p68以及多蛋白黏连蛋白和联会复合体的成分是NPB的必要组成部分。我们的结果支持NPB作为染色中心前体的观点。

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