Hiraoka Y, Dernburg A F, Parmelee S J, Rykowski M C, Agard D A, Sedat J W
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0554.
J Cell Biol. 1993 Feb;120(3):591-600. doi: 10.1083/jcb.120.3.591.
We have determined the position within the nucleus of homologous sites of the histone gene cluster in Drosophila melanogaster using in situ hybridization and high-resolution, three-dimensional wide field fluorescence microscopy. A 4.8-kb biotinylated probe for the histone gene repeat, located approximately midway along the short arm of chromosome 2, was hybridized to whole-mount embryos in late syncytial and early cellular blastoderm stages. Our results show that the two homologous histone loci are distinct and separate through all stages of the cell cycle up to nuclear cycle 13. By dramatic contrast, the two homologous clusters were found to colocalize with high frequency during interphase of cycle 14. Concomitant with homolog pairing at cycle 14, both histone loci were also found to move from their position near the midline of the nucleus toward the apical side. This result suggests that coincident with the initiation of zygotic transcription, there is dramatic chromosome and nuclear reorganization between nuclear cycles 13 and 14.
我们利用原位杂交以及高分辨率三维宽视野荧光显微镜,确定了黑腹果蝇组蛋白基因簇同源位点在细胞核内的位置。用于组蛋白基因重复序列的一个4.8千碱基对的生物素化探针,位于2号染色体短臂大约中间位置,与处于合胞体晚期和细胞胚盘早期的整装胚胎进行杂交。我们的结果表明,在直至核周期13的细胞周期所有阶段,两个同源组蛋白基因座都是不同且分离的。与之形成鲜明对比的是,在周期14的间期,发现两个同源簇高频共定位。伴随着周期14的同源配对,还发现两个组蛋白基因座都从它们在细胞核中线附近的位置向顶端侧移动。这一结果表明,与合子转录的起始同时发生的是,在核周期13和14之间存在显著的染色体和细胞核重组。