Department of Biology, Faculty of Sciences, Kyushu University, Fukuoka 812-8581, Japan.
Am J Bot. 2013 Jan;100(1):60-9. doi: 10.3732/ajb.1200308. Epub 2012 Nov 28.
Phototropins (phot) are blue light receptor proteins that mediate phototropism and control photomovement responses, such as chloroplast photorelocation movement and stomatal opening. Arabidopsis thaliana has two phototropins, phot1 and phot2. Although both phot1 and phot2 redundantly mediate photomovement responses, phot2 uniquely regulates phototropism and the chloroplast avoidance response under high-intensity blue light. However, compared to that of phot1, the mechanistic basis of phot2 function is poorly understood, and in particular, the importance of the LOV2 domain in phot2 function has not been clearly demonstrated. Indeed, photocycle-deficient LOV2 transgenic lines expressing phot2 in a phot1phot2 mutant background retained phototropism, although with less sensitivity than wild-type plants.
We isolated 11 alleles of phot2 mutants and determined the molecular lesion in each allele. We analyzed hypocotyl phototropism, chloroplast photorelocation movement, and leaf flattening in the phot2 mutant and the respective phot1phot2 double mutant plants.
We demonstrated that unlike the phot2 null mutant, the phot2-10 mutant, which has the defective phot2 LOV2 domain, retained the phototropic response and had unusual chloroplast movement. Mutants phot2-2 and phot2-6, which have a missense mutation in the kinase activation loop of phot2, had the phot2-null mutant phenotype. Furthermore, we convincingly demonstrated that the commonly used phot2-1 mutant allele is a phot2-null mutant.
The analyses of the multiple phot2 mutant alleles provided strong evidence for the importance of both LOV domains and the kinase activation loop of phot2 in phototropism and other phot-dependent responses and also demonstrated that phot2-1 allele is a null mutant.
光受体蛋白(phot)是介导向光性和控制光运动反应的蓝光受体蛋白,例如叶绿体光定位运动和气孔开放。拟南芥有两种光受体蛋白,phot1 和 phot2。尽管 phot1 和 phot2 都可以冗余地介导光运动反应,但 phot2 独特地调节高光强下的向光性和叶绿体回避反应。然而,与 phot1 相比,phot2 功能的机制基础了解甚少,特别是 LOV2 结构域在 phot2 功能中的重要性尚未得到明确证明。事实上,在 phot1phot2 突变体背景下表达 phot2 的缺乏光循环 LOV2 结构域的转基因系保留了向光性,尽管敏感性低于野生型植物。
我们分离了 11 个 phot2 突变体等位基因,并确定了每个等位基因的分子病变。我们分析了 phot2 突变体和相应的 phot1phot2 双突变体植物的下胚轴向光性、叶绿体光定位运动和叶片扁平化。
我们证明,与 phot2 无效突变体不同,具有缺陷的 phot2 LOV2 结构域的 phot2-10 突变体保留了向光性反应,并且叶绿体运动异常。phot2-2 和 phot2-6 突变体激酶激活环中有一个错义突变,具有 phot2 无效突变体表型。此外,我们令人信服地证明了常用的 phot2-1 突变体等位基因是一个 phot2 无效突变体。
对多个 phot2 突变体等位基因的分析为 LOV 结构域和 phot2 的激酶激活环在向光性和其他光依赖性反应中的重要性提供了有力证据,同时也证明了 phot2-1 等位基因是一个无效突变体。