Wunsch A M, Lough J
Department of Anatomy and Cellular Biology, Medical College of Wisconsin, Milwaukee 53226.
J Cell Physiol. 1989 Oct;141(1):97-102. doi: 10.1002/jcp.1041410115.
We recently reported that cultures of terminally differentiating myotube cells synthesize histones in reduced but significant amounts in comparison with proliferating myoblasts (Wunsch et al., 1987, Dev. Biol., 119: 85-93). In this study, the stability of myotube histone has been determined, comparing the degradation of de novo-synthesized histones in nascent (day 3) and maturing (day 4) myotubes with histones in the same cells that had been previously made during myoblast proliferation (day 1). Histones synthesized in proliferating myoblasts and myotubes were pulse-labeled with 3H-lysine and chased up to seven days, followed by determinations of radioactivity remaining in histone bands using fluorography of one- and two-dimensional polyacrylamide gels. Considered in aggregate, core histones synthesized de novo in nascent (day 3) myotubes were degraded most rapidly, followed by myotube histones that had been previously made during the proliferative phase (day 1) of myogenesis. De novo-synthesized histones in maturing (day 4) myotubes were relatively stable. Individual histone classes were degraded in the following order of increasing half-life, regardless of the differentiative stage at which they were synthesized: H2A.Z, H2A, H2B, H3(.2, day 1; .3, days 3 and 4), H4.
我们最近报道,与增殖的成肌细胞相比,终末分化的肌管细胞培养物合成组蛋白的量减少但仍很显著(Wunsch等人,1987年,《发育生物学》,119:85 - 93)。在本研究中,已测定了肌管组蛋白的稳定性,将新生(第3天)和成熟(第4天)肌管中从头合成的组蛋白的降解情况与成肌细胞增殖期(第1天)同一细胞中先前合成的组蛋白进行比较。用³H - 赖氨酸对增殖的成肌细胞和肌管中合成的组蛋白进行脉冲标记,并追踪长达7天,然后使用一维和二维聚丙烯酰胺凝胶荧光自显影法测定组蛋白条带中剩余的放射性。总体来看,新生(第3天)肌管中从头合成的核心组蛋白降解最快,其次是在肌生成增殖期(第1天)先前合成的肌管组蛋白。成熟(第4天)肌管中从头合成的组蛋白相对稳定。无论合成时的分化阶段如何,各个组蛋白类别的降解半衰期增加顺序如下:H2A.Z、H2A、H2B、H3(.2,第1天;.3,第3天和第4天)、H4。