Chang Chien-wei, Sun Tai-ping
Department of Biology, Duke University, Durham, NC 27708-1000, USA.
Plant Mol Biol. 2002 Aug;49(6):579-89. doi: 10.1023/a:1015592122142.
The Arabidopsis GA1 gene encodes copalyl diphosphate synthase, which catalyzes the first committed step in the gibberellin biosynthetic pathway. Previous studies indicated that the expression pattern of the GA1 gene is tissue-specific and cell-type-specific during development. Here we showed that expression of GA1 cDNA driven by the 2.4 kb 5'-upstream sequence plus the GA1 genomic coding region into the third exon was able to rescue the gal-3 mutant phenotype. To understand the mechanism controlling GA1 gene expression, cis-regulatory regions in the GA1 promoter were identified by promoter deletion analysis with the GA1-beta-glucuronidase (GUS) gene fusion system. The second intron and the region from -1391 to -997, with respect to the translation initiation site, positively regulate overall GA1-GUS expression level in all tissues examined. Several additional regulatory regions are involved in GA1-GUS expression in all the stages except in seeds: two positive regulatory regions in the first intron and the sequence between -425 and -207, and a negative regulatory region between -1848 and -1391. We also found that the region between -997 and -796 is essential for a high level of GA1 expression in developing seeds.
拟南芥GA1基因编码柯巴基二磷酸合酶,该酶催化赤霉素生物合成途径中的首个关键步骤。先前的研究表明,GA1基因在发育过程中的表达模式具有组织特异性和细胞类型特异性。在此我们表明,由2.4 kb的5'上游序列加上GA1基因组编码区直至第三个外显子驱动的GA1 cDNA表达能够挽救gal-3突变体表型。为了了解控制GA1基因表达的机制,通过使用GA1-β-葡萄糖醛酸酶(GUS)基因融合系统进行启动子缺失分析,确定了GA1启动子中的顺式调控区域。相对于翻译起始位点,第二个内含子以及从-1391至-997的区域对所有检测组织中的总体GA1-GUS表达水平具有正向调控作用。在除种子外的所有阶段,GA1-GUS表达还涉及几个额外的调控区域:第一个内含子中的两个正向调控区域以及-425至-207之间的序列,还有-1848至-1391之间的一个负向调控区域。我们还发现,-997至-796之间的区域对于发育中的种子中高水平的GA1表达至关重要。