Sanchez F, Tobin S L, Rdest U, Zulauf E, McCarthy B J
J Mol Biol. 1983 Feb 5;163(4):533-51. doi: 10.1016/0022-2836(83)90111-0.
DNA fragments representing the six Drosophila actin genes have been isolated by recombinant DNA techniques. We have compared the transcriptional characteristics of the actin genes at the cytological loci 79B and 88F. The activity of each gene in vivo was examined using gene-specific probes from transcribed, but non-translated 3' regions of each gene. The genes show similar patterns of transcriptional activity during development until the pupal stage, with two periods showing RNA accumulation at two to three hours and 12 to 15 hours during embryonic development, followed by large increases in the proportion of message from each gene in first and second instar larvae. During pupal development, the 88F gene apparently produces a larger proportion of transcripts than at any other developmental stage, while the transcripts of the 79B gene are reduced to a level lower than in first and second instar larvae. The 5' end of each messenger RNA in larvae has been mapped by nuclease S1 digestion of hybrids between restriction fragments of genes and homologous mRNAs. The two genes display widely differing capacities to serve as templates for transcription in vitro in HeLa cell extracts. The complete DNA sequences of both genes including the flanking regions immediately 3' and 5' to the gene are presented. These data permit comparison of the DNA sequences of these Drosophila actin genes with each other and with the DNA sequence and protein sequence information available for the actins of Drosophila and other organisms. These two genes share the common structural feature of an intervening sequence at amino acid 307, though the sequences within each intron differ greatly. This may be a reflection of a duplication event, followed by divergence of the intervening sequences. We discuss possible correlations between the DNA sequences of each 5' flanking region and the differences in transcriptional characteristics of these two distinct but closely related genes.
利用重组DNA技术已分离出代表六个果蝇肌动蛋白基因的DNA片段。我们比较了位于79B和88F细胞学位点的肌动蛋白基因的转录特性。使用来自每个基因转录但未翻译的3'区域的基因特异性探针检测了每个基因在体内的活性。这些基因在发育到蛹期之前显示出相似的转录活性模式,在胚胎发育期间有两个时期显示RNA积累,分别在两到三小时和十二到十五小时,随后在一龄和二龄幼虫中每个基因的信使RNA比例大幅增加。在蛹发育期间,88F基因显然产生的转录本比例比任何其他发育阶段都要大,而79B基因的转录本则减少到低于一龄和二龄幼虫的水平。通过对基因的限制性片段与同源mRNA之间的杂交体进行核酸酶S1消化,绘制了幼虫中每个信使RNA的5'末端图谱。这两个基因在HeLa细胞提取物中作为体外转录模板的能力差异很大。给出了这两个基因包括基因两侧紧邻的3'和5'区域的完整DNA序列。这些数据允许将这些果蝇肌动蛋白基因的DNA序列相互比较,并与果蝇和其他生物的肌动蛋白的DNA序列和蛋白质序列信息进行比较。这两个基因在氨基酸307处具有内含子序列这一共同结构特征,尽管每个内含子内的序列差异很大。这可能反映了一次重复事件,随后内含子序列发生了分歧。我们讨论了每个5'侧翼区域的DNA序列与这两个不同但密切相关的基因转录特性差异之间可能的相关性。