DeSalle R, Slightom J, Zimmer E
Genetics. 1986 Apr;112(4):861-75. doi: 10.1093/genetics/112.4.861.
Restriction endonuclease cleavage analyses of cloned and genomic DNA samples indicate that the structure of the DNA encoding the large cytoplasmic RNAs (rDNAs) is altered in Drosophila mercatorum lines which exhibit an abnormal abdomen (aa) phenotype. In a majority of the rDNA repeat units from aa flies, the 28S coding sequence is interrupted by a large [5-6 kilobase pairs (kbp)] insert. A subclone containing this inserted DNA (ins 3) hybridizes primarily to rDNA-containing sequences in in situ and genomic blot hybridization experiments. Additionally, genomic nitrocellulose blot hybridization analyses show that ins- containing rDNA repeat units are clustered in a spontaneously arising aa mutant. This rDNA alteration in D. mercatorum flies with the aa phenotype more closely resembles the bobbed (bb) defect of D. hydei than the bb defect of D. melanogaster, which involves alterations in rDNA copy number. By analogy with the other Drosophila systems, we propose that the altered D. mercatorum rDNA repeat units are defective in rRNA production at a critical stage. The lowered levels of rRNA ultimately would limit the concentration of ribosomes needed to produce large quantities of a protein (in these cases, juvenile hormone esterase) needed for normal development.
对克隆的和基因组DNA样本进行的限制性内切酶切割分析表明,在表现出异常腹部(aa)表型的黑腹果蝇品系中,编码大细胞质RNA(rDNAs)的DNA结构发生了改变。在大多数来自aa果蝇的rDNA重复单元中,28S编码序列被一个大的[5 - 6千碱基对(kbp)]插入片段打断。在原位杂交和基因组印迹杂交实验中,一个包含该插入DNA(ins 3)的亚克隆主要与含rDNA的序列杂交。此外,基因组硝酸纤维素印迹杂交分析表明,含有ins的rDNA重复单元在一个自发产生的aa突变体中聚集。具有aa表型的黑腹果蝇的这种rDNA改变与海德氏果蝇的截毛(bb)缺陷比与黑腹果蝇的bb缺陷更为相似,黑腹果蝇的bb缺陷涉及rDNA拷贝数的改变。通过与其他果蝇系统类比,我们提出,改变后的黑腹果蝇rDNA重复单元在rRNA产生的关键阶段存在缺陷。rRNA水平的降低最终会限制产生正常发育所需大量蛋白质(在这些情况下是保幼激素酯酶)所需核糖体的浓度。