Boylan M, Douglas N, Quail P H
University of California-Berkeley/U.S. Department of Agriculture, Plant Gene Expression Center, Albany 94710.
Plant Cell. 1994 Mar;6(3):449-60. doi: 10.1105/tpc.6.3.449.
We used the exaggerated short hypocotyl phenotype induced by oat phytochrome A overexpression in transgenic Arabidopsis to monitor the biological activity of mutant phytochrome A derivatives. Three different mutations, which were generated by removing 52 amino acids from the N terminus (delta N52), the entire C-terminal domain (delta C617), or amino acids 617-686 (delta 617-686) of the oat molecule, each caused striking dominant negative interference with the ability of endogenous Arabidopsis phytochrome A to inhibit hypocotyl growth in continuous far-red light ("far-red high irradiance response" conditions). By contrast, in continuous white or red light, delta N52 was as active as the unmutagenized oat phytochrome A protein in suppressing hypocotyl elongation, while delta C617 and delta 617-686 continued to exhibit dominant negative behavior under these conditions. These data suggest that at least three spatially discrete molecular domains coordinate the photoregulatory activities of phytochrome A in Arabidopsis seedlings. The first is the chromophore-bearing N-terminal domain between residues 53 and 616 that is apparently sufficient for the light-induced initiation but not the completion of productive interactions with transduction chain components. The second is the C-terminal domain between residues 617 and 1129 that is apparently necessary for completion of productive interactions under all irradiation conditions. The third is the N-terminal 52 amino acids that are apparently necessary for completion of productive interactions only under far-red high irradiance conditions and are completely dispensable under white and red light regimes.
我们利用燕麦光敏色素A在转基因拟南芥中过表达所诱导的短下胚轴表型来监测突变型光敏色素A衍生物的生物活性。通过去除燕麦分子N端的52个氨基酸(δN52)、整个C端结构域(δC617)或氨基酸617 - 686(δ617 - 686)产生了三种不同的突变,每种突变都对拟南芥内源性光敏色素A在持续远红光下抑制下胚轴生长的能力(“远红光高辐照反应”条件)产生了显著的显性负干扰。相比之下,在持续白光或红光下,δN52在抑制下胚轴伸长方面与未诱变的燕麦光敏色素A蛋白活性相当,而δC617和δ617 - 686在这些条件下仍表现出显性负行为。这些数据表明,至少三个空间上离散的分子结构域协调了拟南芥幼苗中光敏色素A的光调节活性。第一个是残基53至616之间带有生色团的N端结构域,它显然足以引发光诱导的起始,但不足以完成与转导链成分的有效相互作用。第二个是残基617至1129之间的C端结构域,它显然是在所有辐照条件下完成有效相互作用所必需的。第三个是N端的52个氨基酸,它们显然仅在远红光高辐照条件下对于完成有效相互作用是必需的,而在白光和红光条件下则完全 dispensable(此处原文有误,应是“dispensable”,意为“可有可无的”) 。