Mason H S, DeWald D B, Mullet J E
Department of Biochemistry and Biophysics, Texas A&M University, College Station 77843-2128.
Plant Cell. 1993 Mar;5(3):241-51. doi: 10.1105/tpc.5.3.241.
Soybean vspB encodes a highly expressed vegetative storage protein-acid phosphatase. In soybean, vspB expression is stimulated by methyl jasmonate (MeJA) and sugars. The vspB promoter was studied by transforming tobacco with fusions of 5' noncoding vspB DNA and the gene encoding beta-glucuronidase (GUS). Constructs containing 833 bp of vspB 5' DNA showed high expression of GUS in stems, leaf veins and trichomes, sepals, and pollen. Sucrose (0.2 M) and MeJA (10(-5) M) increased gene expression when applied to leaf tissue. Deletion of the region -787 to -520 with respect to the transcription initiation site rendered the vspB promoter noninducible by MeJA but still sucrose responsive. This result indicates that DNA elements capable of modulating vspB by MeJA can be separated from carbon response elements. Further 5' end deletion from -520 to -403 or 3' end deletion from -165 to -289 removed DNA sequences involved in carbon modulation of gene expression. A DNA domain that mediates the MeJA response was further localized to a 50-bp region between -535 and -585. This domain when fused to a cauliflower mosaic virus (CaMV) 35S truncated (-88) promoter makes the CaMV promoter responsive to MeJA. The MeJA-responsive domain contains a G-box motif (CACGTG) and a C-rich sequence. A similar 50-bp DNA region is present in the putative promoter of vspA. Related sequences are located in a wound- and MeJA-responsive domain of the proteinase inhibitor II gene and a UV-responsive promoter domain of chs, the gene encoding chalcone synthase that is also responsive to MeJA.
大豆vspB编码一种高表达的营养贮藏蛋白-酸性磷酸酶。在大豆中,vspB的表达受茉莉酸甲酯(MeJA)和糖类的刺激。通过用5'非编码vspB DNA与编码β-葡萄糖醛酸酶(GUS)的基因的融合体转化烟草来研究vspB启动子。含有833 bp的vspB 5' DNA的构建体在茎、叶脉和毛状体、萼片以及花粉中显示出GUS的高表达。当应用于叶片组织时,蔗糖(0.2 M)和MeJA(10^(-5) M)可增加基因表达。相对于转录起始位点缺失-787至-520区域,使vspB启动子对MeJA无诱导性,但仍对蔗糖有反应。该结果表明,能够被MeJA调节vspB的DNA元件可与碳反应元件分离。从-520至-403的进一步5'端缺失或从-165至-289的3'端缺失去除了参与基因表达碳调节的DNA序列。介导MeJA反应的一个DNA结构域进一步定位于-535至-585之间的一个50 bp区域。该结构域与花椰菜花叶病毒(CaMV)35S截短(-88)启动子融合后,使CaMV启动子对MeJA有反应。MeJA反应结构域包含一个G-box基序(CACGTG)和一个富含C的序列。在vspA的推定启动子中存在一个类似的50 bp DNA区域。相关序列位于蛋白酶抑制剂II基因的伤口和MeJA反应结构域以及chs(编码查尔酮合酶且也对MeJA有反应的基因)的紫外线反应启动子结构域中。