Muramoto T, Kohchi T, Yokota A, Hwang I, Goodman H M
Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan.
Plant Cell. 1999 Mar;11(3):335-48. doi: 10.1105/tpc.11.3.335.
The HY1 locus of Arabidopsis is necessary for phytochrome chromophore biosynthesis and is defined by mutants that show a long hypocotyl phenotype when grown in the light. We describe here the molecular cloning of the HY1 gene by using chromosome walking and mutant complementation. The product of the HY1 gene shows significant similarity to animal heme oxygenases and contains a possible transit peptide for transport to plastids. Heme oxygenase activity was detected in the HY1 protein expressed in Escherichia coli. Heme oxygenase catalyzes the oxygenation of heme to biliverdin, an activity that is necessary for phytochrome chromophore biosynthesis. The predicted transit peptide is sufficient to transport the green fluorescent protein into chloroplasts. The accumulation of the HY1 protein in plastids was detected by using immunoblot analysis with an anti-HY1 antiserum. These results indicate that the Arabidopsis HY1 gene encodes a plastid heme oxygenase necessary for phytochrome chromophore biosynthesis.
拟南芥的HY1基因座对于光敏色素生色团的生物合成是必需的,其由在光照下生长时表现出长下胚轴表型的突变体所定义。我们在此描述了通过染色体步移和突变体互补对HY1基因进行分子克隆的过程。HY1基因的产物与动物血红素加氧酶具有显著的相似性,并含有一个可能用于转运至质体的转运肽。在大肠杆菌中表达的HY1蛋白中检测到了血红素加氧酶活性。血红素加氧酶催化血红素氧化为胆绿素,这一活性对于光敏色素生色团的生物合成是必需的。预测的转运肽足以将绿色荧光蛋白转运至叶绿体。通过使用抗HY1抗血清的免疫印迹分析检测到了HY1蛋白在质体中的积累。这些结果表明,拟南芥HY1基因编码一种对于光敏色素生色团生物合成必需的质体血红素加氧酶。