Couderc J L, Hilal L, Sobrier M L, Dastugue B
Nucleic Acids Res. 1987 Mar 25;15(6):2549-61. doi: 10.1093/nar/15.6.2549.
The steroid hormone 20-hydroxyecdysone (20-OHE) induces, in Kc cultured Drosophila melanogaster cells, important morphological transformations and specific changes of enzymatic activities and of protein synthesis. These changes are accompanied by an increase of synthesis and an accumulation of actin. Specific probes were used to reveal transcripts of each actin gene in mRNA populations isolated from cells at various times of 20-OHE treatment. Only the two cytoplasmic actin genes 5C and 42A are expressed in Kc cells and the hormone induces the accumulation of transcripts of these two genes. We have also taken advantage of S1 mapping and extension procedures to identify the 5' ends of the actin mRNAs from these two genes and to compare their respective levels of expression. The 5C gene is more expressed than the 42A one in untreated and in hormone treated cells. The 5C gene encodes three RNAs that differ in their 3' end. The two genes are interrupted by an intervening sequence immediately upstream of ATG initiation codon but not at the same position. The transcription rate for the two genes is increased up to five fold upon 20-OHE treatment, demonstrating a direct effect of the steroid hormone at the transcriptional level for these genes.
类固醇激素20-羟基蜕皮酮(20-OHE)在培养的果蝇Kc细胞中可诱导重要的形态转变以及酶活性和蛋白质合成的特定变化。这些变化伴随着肌动蛋白合成的增加和积累。使用特异性探针来揭示在20-OHE处理的不同时间从细胞中分离的mRNA群体中每个肌动蛋白基因的转录本。在Kc细胞中仅表达两个细胞质肌动蛋白基因5C和42A,并且该激素诱导这两个基因转录本的积累。我们还利用S1作图和延伸程序来鉴定来自这两个基因的肌动蛋白mRNA的5'末端,并比较它们各自的表达水平。在未处理和激素处理的细胞中,5C基因的表达均高于42A基因。5C基因编码三种3'末端不同的RNA。这两个基因在ATG起始密码子上游紧邻处被一个间隔序列中断,但位置不同。经20-OHE处理后,这两个基因的转录速率提高了五倍,表明类固醇激素对这些基因在转录水平上具有直接作用。