Kellum R, Raff J W, Alberts B M
Department of Biochemistry and Biophysics, University of California at San Francisco 94143-0448, USA.
J Cell Sci. 1995 Apr;108 ( Pt 4):1407-18. doi: 10.1242/jcs.108.4.1407.
Heterochromatin protein 1 (HP1) was initially discovered as a protein that is associated with the heterochromatin at the chromocenter of polytene chromosomes in Drosophila larval salivary glands. In this paper we investigate the localization of heterochromatin protein 1 in the diploid nuclei of Drosophila embryos. We focus on its association with the interphase heterochromatin in fixed embryos before and during cycle 14, the developmental time at which heterochromatin becomes most conspicuous, and also follow its localization during mitosis. The GAGA transcription factor was recently shown to be localized at sequences within alpha-heterochromatin in pre-cycle 14 embryos, and an antibody against this protein serves as a convenient marker for these sequences. We find an enrichment of heterochromatin protein 1 in the intensely DAPI-staining regions near the apical surface of nuclear cycle 10 embryos. At this stage GAGA factor is localized into punctate structures in this same region. This enrichment for HP1 is markedly increased during nuclear cycle 14. Surprisingly, whereas GAGA factor retains its association with the heterochromatin throughout the cell cycle, a significant fraction of HP1 is dispersed throughout the spindle around the segregating chromosomes during mitosis. This dispersed pool of heterochromatin protein 1 was observed during mitosis in both early and late Drosophila embryos and in an analysis of a bacterially produced 6x histidine-heterochromatin protein 1 fusion protein injected into living Drosophila embryos. When Drosophila tissue culture cells were prepared by a method which removes soluble protein and avoids fixation of the mitotic chromosomes, an enrichment for heterochromatin protein 1 in the heterochromatin of the chromosomes was discovered also.
异染色质蛋白1(HP1)最初是作为一种与果蝇幼虫唾液腺多线染色体着丝粒处的异染色质相关的蛋白质被发现的。在本文中,我们研究了异染色质蛋白1在果蝇胚胎二倍体细胞核中的定位。我们关注其在第14个细胞周期之前和期间固定胚胎中的间期异染色质的关联,这是异染色质变得最为明显的发育阶段,同时也追踪其在有丝分裂期间的定位。最近发现GAGA转录因子定位于第14个细胞周期之前胚胎的α-异染色质内的序列,并且针对该蛋白质的抗体可作为这些序列的便捷标记。我们发现在核周期10胚胎顶端表面附近强烈DAPI染色区域中异染色质蛋白1富集。在此阶段,GAGA因子定位于同一区域的点状结构中。在核周期14期间,HP1的这种富集显著增加。令人惊讶的是,尽管GAGA因子在整个细胞周期中都与异染色质保持关联,但在有丝分裂期间,相当一部分HP1分散在分离染色体周围的纺锤体中。在早期和晚期果蝇胚胎的有丝分裂过程中以及对注入活果蝇胚胎中的细菌产生的6x组氨酸 - 异染色质蛋白1融合蛋白的分析中都观察到了这种分散的异染色质蛋白1池。当通过去除可溶性蛋白并避免有丝分裂染色体固定的方法制备果蝇组织培养细胞时,也发现染色体异染色质中异染色质蛋白1富集。