Rudert F, Bronner S, Garnier J M, Dollé P
Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, INSERM, IGBMC, Illkirch, France.
Mamm Genome. 1995 Feb;6(2):76-83. doi: 10.1007/BF00303248.
Using in vitro immuno-selected retinoic acid response elements, we have isolated mouse genomic clones containing major (gamma) satellite DNA repeats that are considered as typical of chromosome centromeres. Several cDNA clones were then isolated from a F9 cell cDNA library and were found to harbor variants of the 234-base pair consensus gamma satellite monomer. In Northern analysis, these satellite DNA sequences hybridized predominantly to an approximately 1.8-kb RNA species in polyadenylated RNA from P19 cells. These transcripts were strongly repressed by retinoic acid, and nuclear run-on assays revealed that this repression was, at least in part, mediated at the transcriptional level. Satellite transcripts were also detected in HeLa cells, where they were similarly down-regulated by retinoids. Heterogeneously sized satellite transcripts were detected in RNA from specific mouse tissues, such as fetuses (but not placenta), adult liver, and testis. In situ hybridization analysis revealed that satellite transcripts are generated from opposite DNA strands and are differentially expressed in cells of the developing central nervous system as well as in adult liver and testis. These data may have implications on retinoic acid-mediated transcriptional regulation and centromere function.
利用体外免疫选择的视黄酸反应元件，我们分离出了包含主要（γ）卫星DNA重复序列的小鼠基因组克隆，这些重复序列被认为是染色体着丝粒的典型特征。随后从F9细胞cDNA文库中分离出几个cDNA克隆，发现它们含有234个碱基对的共有γ卫星单体的变体。在Northern分析中，这些卫星DNA序列主要与来自P19细胞的多聚腺苷酸化RNA中的一种约1.8kb的RNA物种杂交。这些转录本被视黄酸强烈抑制，核转录分析表明这种抑制至少部分是在转录水平介导的。在HeLa细胞中也检测到了卫星转录本，在那里它们同样被类视黄醇下调。在来自特定小鼠组织（如胎儿（但不包括胎盘）、成年肝脏和睾丸）的RNA中检测到了大小各异的卫星转录本。原位杂交分析表明，卫星转录本由相反的DNA链产生，并在发育中的中枢神经系统细胞以及成年肝脏和睾丸中差异表达。这些数据可能对视黄酸介导的转录调控和着丝粒功能有影响。
