Botany Department, University of Wisconsin, Madison, Wisconsin 53706.
Proc Natl Acad Sci U S A. 1983 Apr;80(8):2248-52. doi: 10.1073/pnas.80.8.2248.
Translatable phytochrome mRNA represents approximately 5 x 10(-3)% of the total poly(A)-RNA present in etiolated Avena seedlings, as determined by incorporation of radioactivity into the immunoprecipitable apoprotein in a cell-free translation system. Irradiation of such seedlings with 5-s red light induces a decline in this mRNA that is detectable within 15-30 min, shows a 50% reduction within 50-60 min, and results in a >95% reduction within 2 hr. The effect of the red light pulse is reversed by an immediately subsequent far-red pulse to the level of the far-red-light control, indicating that phytochrome exerts autoregulatory control over its own translatable mRNA level. This result necessitates revision of existing concepts of how phytochrome concentrations are modulated in vivo. Red-light dose-response curves show that the response is sensitive to very low light levels. Conversion of <1% of the total cellular phytochrome to the biologically active far-red-absorbing form is sufficient to induce approximately 60% of the maximal response, and 20% far-red-absorbing form saturates the response. The observed change in translatable phytochrome mRNA level is one of the most rapid phytochrome-induced alterations in any cellular mRNA yet recorded. Thus, autoregulation of phytochrome mRNA provides an attractive opportunity to examine the early sequence of events in phytochrome control of gene expression.
可翻译的光敏色素 mRNA 约占暗培养的燕麦幼苗中总 poly(A)-RNA 的 5 x 10(-3)%,这是通过在无细胞翻译系统中掺入放射性物质到免疫沉淀的脱辅基蛋白来确定的。用 5-s 红光照射这些幼苗会导致这种 mRNA 的下降,在 15-30 分钟内即可检测到,在 50-60 分钟内减少 50%,在 2 小时内减少 >95%。红光脉冲的作用可被随后的远红光脉冲逆转至远红光对照的水平,表明光敏色素对其自身可翻译的 mRNA 水平进行自身调节控制。这一结果需要对体内如何调节光敏色素浓度的现有概念进行修正。红光剂量反应曲线表明,该反应对非常低的光水平敏感。将总细胞光敏色素的 <1%转化为生物活性的远红光吸收形式足以诱导约 60%的最大反应,而 20%的远红光吸收形式使反应饱和。可翻译的光敏色素 mRNA 水平的观察到的变化是迄今为止在任何细胞 mRNA 中记录到的最快的光敏色素诱导变化之一。因此,光敏色素 mRNA 的自身调节为研究光敏色素对基因表达的控制的早期事件提供了一个有吸引力的机会。