Pipkin J, Anson J, Hinson W, Hudson J
Biochim Biophys Acta. 1982 Nov 30;699(2):155-63. doi: 10.1016/0167-4781(82)90149-x.
The in vivo administration of hydroxyurea for 12 h counteracts DNA synthesis and cell cycling stimulated by 72 h of isoproterenol treatment in rat salivary gland, as determined by fluorescence-activated flow cytometry. Hydroxyurea has little effect on [3H]leucine incorporation (protein synthesis) of the nuclear proteins soluble in 0.35 M NaCl, when examined by polyacrylamide gel chromatography and autoradiography from electrostatically sorted nuclei of (G0 + G1) and (G2 + M) phases of the in vivo cell cycle. Differential incorporation of [3H]leucine into nuclear proteins was observed during various phases of the cell cycle. Proteins 'X' and 'Z', observed in stained gel chromatographs of the 0.35 M NaCl-soluble nuclear proteins, were identified by biochemical analyses as ubiquitin and protein A24, respectively. Ubiquitin appeared transiently while A24 increased in gel chromatograms concomitant with progressive quiescence of the salivary gland induced by hydroxyurea.
通过荧光激活流式细胞术测定,在大鼠唾液腺中,体内给予羟基脲12小时可抵消异丙肾上腺素处理72小时所刺激的DNA合成和细胞周期进程。当通过聚丙烯酰胺凝胶色谱法和放射自显影术,对体内细胞周期(G0 + G1)期和(G2 + M)期经静电分选的细胞核中可溶于0.35 M NaCl的核蛋白的[3H]亮氨酸掺入(蛋白质合成)进行检测时,羟基脲对其影响很小。在细胞周期的各个阶段观察到[3H]亮氨酸在核蛋白中的差异掺入。在0.35 M NaCl可溶核蛋白的染色凝胶色谱图中观察到的蛋白质“X”和“Z”,经生化分析分别鉴定为泛素和蛋白质A24。泛素短暂出现,而在凝胶色谱图中,随着羟基脲诱导唾液腺逐渐静止,A24增加。