Clark S H, Kiefer B I
Genetics. 1977 Aug;86(4):801-11. doi: 10.1093/genetics/86.4.801.
It has been demonstrated that a particular rDNA-deficient Y chromosome (Y(bbSuVar-5)) increases the rate of ribosomal RNA synthesis in adult testes (Shermoen and Kiefer 1975) and in whole flies (Clark, Strausbaugh and Kiefer 1977). As an initial attempt to explore the molecular basis of this phenomenon, experiments were designed to determine if the rate increase was specific for rRNA as opposed to the other species of RNA. The genotypes used in these studies were car bb/Y(bb-), car bb/Y( bbSuVar-5), and Sam(+) iso. car bb/Y(bbSuVar-5 ) and car bb/Y(bb-) are deficient to the same extent in rDNA and Sam(+) iso is a wild-type stock. Following isotope incorporation, total RNA was extracted by a phenol:chloroform method and separated by polyacrylamide gel electrophoresis. The various RNA species were quantified by UV absorption and their radioactivity determined by gel fractionation and liquid scintillation counting. The resulting data permitted the calculation of a specific activity (i.e., dpm/microg RNA) which was defined as synthetic rate. Polyadenylated RNA was isolated using a poly-U sepharose column and similar rate calculations were made. The data from these studies indicate that the rate of synthesis of all species of RNA examined (28S + 18S, 5S, 4S transfer RNA and polyadenylated RNA) is increased by the presence of the Y(bbSuVar-5) chromosome. Genetic and molecular mechanisms are discussed.
已经证明,一种特定的rDNA缺陷型Y染色体(Y(bbSuVar - 5))会提高成年睾丸(Shermoen和Kiefer,1975年)以及整个果蝇(Clark、Strausbaugh和Kiefer,1977年)中核糖体RNA的合成速率。作为探索这一现象分子基础的初步尝试,设计了实验来确定合成速率的增加是否对rRNA具有特异性,而非针对其他种类的RNA。这些研究中使用的基因型为car bb/Y(bb -)、car bb/Y(bbSuVar - 5)和Sam(+) iso。car bb/Y(bbSuVar - 5)和car bb/Y(bb -)在rDNA方面的缺陷程度相同,而Sam(+) iso是野生型品系。在进行同位素掺入后,通过苯酚:氯仿法提取总RNA,并通过聚丙烯酰胺凝胶电泳进行分离。通过紫外吸收对各种RNA种类进行定量,并通过凝胶分级分离和液体闪烁计数确定其放射性。所得数据允许计算比活性(即dpm/μg RNA),其被定义为合成速率。使用聚-U琼脂糖柱分离聚腺苷酸化RNA,并进行类似的速率计算。这些研究的数据表明,Y(bbSuVar - 5)染色体的存在会提高所检测的所有RNA种类(28S + 18S、5S、4S转移RNA和聚腺苷酸化RNA)的合成速率。文中讨论了遗传和分子机制。