Diaz M O, Gall J G
Chromosoma. 1985;92(4):243-53. doi: 10.1007/BF00329807.
We have studied transcription at the histone loci in oocytes of the newt Notophthalmus viridescens, using in situ hybridization of cloned probes to the nascent RNA on lampbrush chromosome loops. Clusters of the five histone coding sequences are separated by long tracts of a simple sequence DNA, satellite 1. We had previously demonstrated coordinate transcription of histone genes and satellite 1 sequences. We postulated that satellite sequences were transcribed by readthrough from histone gene promoters; that is, transcription initiated at any of the five usual promoters, but did not terminate at the 3' end of the gene. Instead transcription proceeded through downstream sequences in the histone cluster (including spacers and downstream histone genes), and then through the satellite 1 region. Our model led to several specific predictions, in particular that some internal spacer regions between the genes should be well represented in the RNA on loops, that certain sequences should be absent from the loops, and that presence or absence of particular sequences should be correlated with morphological polarity of the transcription unit. We have hybridized ten strand-specific probes to the lamp-brush chromosomes and we find that the patterns of hybridization agree with the readthrough model of transcription.
我们利用克隆探针与灯刷染色体环上的新生RNA进行原位杂交,研究了蝾螈绿红东美螈卵母细胞中组蛋白基因座的转录情况。五个组蛋白编码序列簇被长段的简单序列DNA(卫星1)隔开。我们之前已经证明组蛋白基因和卫星1序列是协同转录的。我们推测卫星序列是通过从组蛋白基因启动子通读转录而来的;也就是说,转录在五个常见启动子中的任何一个处起始,但不在基因的3'端终止。相反,转录会继续通过组蛋白簇中的下游序列(包括间隔区和下游组蛋白基因),然后通过卫星1区域。我们的模型产生了几个具体的预测,特别是基因之间的一些内部间隔区在环上的RNA中应该有很好的呈现,环上应该不存在某些序列,并且特定序列的存在或缺失应该与转录单位的形态极性相关。我们已经将十个链特异性探针与灯刷染色体杂交,发现杂交模式与通读转录模型相符。