Leibovich B A, Biliaeva E S, Zhimulev I F, Khesin R B
Ontogenez. 1974;5(6):544-56.
The incorporation of 3H-uridine in different regions of polytene chromosomes in live cells of the Drosophila melanogaster salivary glands was compared with the incorporation of 3H-UTP in the same regions under the incubation of cytological preparations of these chromosomes with the E. coli RNA polymerase. The label distribution by regions was compared with the DNA content in them. Individual regions of chromosomes differ by 3H-uridine incorporation in live cells to a much greater extent than by 3H-UTP incorporation in vitro under the incubation with a non-homologous enzyme. RNA synthesis in an exogenous enzyme depends on the DNA content in different chromosome regions to a much greater extent than RNA synthesis in vivo. The correlation of label distribution after 3H-uridine incorporation in live cells and after RNA synthesis in vitro on the preparations by the bacterial RNA polymerase is, correspondingly, very low. This enzyme forms, however, RNA's on puffs 2-3 times more actively than on the same regions in non-puffing state but this difference is dozens of times greater in live cells. RNA synthesis in vitro is, thus, non-specific and does not correspond practically to the intensity of RNA synthesis on the same chromosome regions in live cells. At the same time, as in live cells, the E. coli enzyme synthesizes twice more RNA on the single X-chromosome of males (1X2A) than on each of X-chromosomes of diploid (2X2A) and triploid (3X3A) females or superfemales (3X2A), whereas in intersexes (2X3A) X-chromosomes display intermediate template activity. Thus, RNA synthesis by a heterologous enzyme in vitro does not differ by this index from the synthesis in live cells. It is suggested that differences in the template activity of X-chromosomes in vitro depending on the sex index (X : A) are due to different degree of DNP condensation in these chromosomes. In spite of differences in the degree of condensation, the male X-chromosome binds on the fixed preparation approximately the same amount of thymus histone F1 carrying fluorochrome as each of two female X-chromosomes. Hence, there is no sharp difference between the male and female X-chromosomes by the number and length of DNA regions accessible for interaction with exogenous proteins. On the basis of the data obtained, a hypothesis about two levels and, respectively, two mechanisms of control gene activity in animal chromosomes is considered. The first mechanism is, supposedly, based on decondensation of DNP appears to result in that the same proteins-regulators in the same amount activate corresponding genes in X-chromosome in males twice more strongly than in females.
比较了3H-尿苷在黑腹果蝇唾液腺活细胞多线染色体不同区域的掺入情况,以及在这些染色体的细胞学制剂与大肠杆菌RNA聚合酶孵育时,3H-UTP在相同区域的掺入情况。将各区域的标记分布与其中的DNA含量进行了比较。染色体的各个区域在活细胞中3H-尿苷掺入的差异程度,比在与非同源酶孵育的体外条件下3H-UTP掺入的差异程度大得多。外源酶催化的RNA合成比体内RNA合成在更大程度上依赖于不同染色体区域的DNA含量。相应地,活细胞中3H-尿苷掺入后与细菌RNA聚合酶在制剂上体外RNA合成后的标记分布相关性非常低。然而,这种酶在胀泡上形成RNA的活性比在非胀泡状态的相同区域高2 - 3倍,但在活细胞中这种差异要大几十倍。因此,体外RNA合成是非特异性的,实际上与活细胞中相同染色体区域的RNA合成强度不相符。同时,与活细胞一样,大肠杆菌酶在雄性单条X染色体(1X2A)上合成的RNA比二倍体(2X2A)和三倍体(3X3A)雌性或超雌体(3X2A)的每条X染色体上合成的RNA多两倍,而在间性体(2X3A)中X染色体表现出中等模板活性。因此,体外异源酶催化的RNA合成在该指标上与活细胞中的合成没有差异。有人认为,体外X染色体模板活性因性指数(X : A)不同而产生差异,是由于这些染色体中DNP凝聚程度不同。尽管凝聚程度存在差异,但在固定制剂上,雄性X染色体结合携带荧光染料的胸腺组蛋白F1的量与两条雌性X染色体中的每一条大致相同。因此,在可与外源蛋白相互作用的DNA区域数量和长度方面,雄性和雌性X染色体之间没有明显差异。基于所获得的数据,考虑了关于动物染色体中基因活性控制的两个水平以及相应的两种机制的假说。第一种机制据推测是基于DNP的解聚,这似乎导致相同数量的相同蛋白质调节因子在雄性X染色体中激活相应基因的强度比在雌性中强两倍。