hp1γ与哺乳动物细胞核和染色体中的常染色质和异染色质有关。

E Minc, J C Courvalin, B Buendia
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引用次数: 174

摘要

异染色质蛋白1 (HP1)是一种非组蛋白染色体蛋白,首次在果蝇中发现,在基因沉默中起剂量依赖性作用。在哺乳动物中有三个同源基因,hp1α, hp1β和hp1γ。虽然hp1α和hp1β通过免疫细胞化学方法明确定位于异染色质,但hp1γ被发现要么只与常染色质相关,要么同时存在于常染色质和异染色质中。在这里,我们使用一种针对分子羧基末端肽表位的抗体,将hp1γ定位在间期细胞核的常染色质和异染色质室中,以及3T3细胞有丝分裂染色体的周中心点染色质和臂中。在与绿色荧光蛋白融合表达hp1γ的细胞核中也观察到这种双重定位。相反,当用针对氨基末端表位的抗体分析hp1γ的分布时,该蛋白在常染色质中检测到,而在异染色质中检测不到,除了在S期后期异染色质进行复制时的短暂异染色质染色。这些数据表明,有争议的hp1γ在染色质中的免疫定位是由于针对拓扑不同的表位的抗体的使用,这些表位存在于分子的氨基末端,在非复制性异染色质中被选择性地掩盖。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
HP1gamma associates with euchromatin and heterochromatin in mammalian nuclei and chromosomes.

Heterochromatin protein 1 (HP1) is a nonhistone chromosomal protein, first identified in Drosophila, that plays a dose-dependent role in gene silencing. Three orthologs, HP1alpha, HP1beta, and HP1gamma, have been characterized in mammals. While HP1alpha and HP1beta have been unambiguously localized in heterochromatin by immunocytochemical methods, HP1gamma has been found either exclusively associated with euchromatin or present in both euchromatin and heterochromatin. Here, using an antibody directed against a peptide epitope at the carboxyl-terminal end of the molecule, we localize HP1gamma in both euchromatin and heterochromatin compartments of interphase nuclei, as well as in the pericentromeric chromatin and arms of mitotic chromosomes of 3T3 cells. This dual location was also observed in nuclei expressing HP1gamma as a fusion protein with green fluorescent protein. In contrast, when the distribution of HP1gamma was analyzed with antibodies directed against an amino-terminal epitope, the protein was detectable in euchromatin and not in heterochromatin, except for transient heterochromatin staining during the late S phase, when the heterochromatin undergoes replication. These data suggest that the controversial immunolocalization of HP1gamma in chromatin is due to the use of antibodies directed against topologically distinct epitopes, those present at the amino-terminal end of the molecule being selectively masked in nonreplicative heterochromatin.

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