Rushton P J, Torres J T, Parniske M, Wernert P, Hahlbrock K, Somssich I E
Max-Planck-Institut für Züchtungsforschung, Köln, Germany.
EMBO J. 1996 Oct 15;15(20):5690-700.
PR1 is a pathogenesis-related protein encoded in the parsley genome by a family of three genes (PR1-1, PR1-2 and PR1-3). Loss- and gain-of-function experiments in a transient expression system demonstrated the presence of two fungal elicitor responsive elements in each of the PR1-1 and PR1-2 promoters. These elements, W1, W2 and W3, contain the sequence (T)TGAC(C) and mutations that disrupt this sequence abolish function. Gel shift experiments demonstrated that W1, W2 and W3 are bound specifically by similar nuclear proteins. Three cDNA clones encoding sequence-specific DNA-binding proteins were isolated by South-Western screening and these proteins, designated WRKY1, 2 and 3, also bind specifically to W1, W2 and W3. WRKY1, 2 and 3 are members of the family of sequence-specific DNA-binding proteins, which we call the WRKY family. Treatment of parsley cells with the specific oligopeptide elicitor Pep25 induced a transient and extremely rapid increase in mRNA levels of WRKY1 and 3. WRKY2 mRNA levels in contrast showed a concomitant transient decrease. These rapid changes in WRKY mRNA levels in response to a defined signal molecule suggest that WRKY1, 2 and 3 play a key role in a signal transduction pathway that leads from elicitor perception to PR1 gene activation.
PR1是一种病程相关蛋白，由欧芹基因组中的三个基因家族（PR1-1、PR1-2和PR1-3）编码。在瞬时表达系统中进行的功能缺失和功能获得实验表明，PR1-1和PR1-2启动子中各自存在两个真菌激发子响应元件。这些元件W1、W2和W3包含序列（T）TGAC（C），破坏该序列的突变会导致功能丧失。凝胶迁移实验表明，W1、W2和W3被相似的核蛋白特异性结合。通过South-Western筛选分离出三个编码序列特异性DNA结合蛋白的cDNA克隆，这些蛋白分别命名为WRKY1、2和3，它们也特异性结合W1、W2和W3。WRKY1、2和3是序列特异性DNA结合蛋白家族的成员，我们将其称为WRKY家族。用特异性寡肽激发子Pep25处理欧芹细胞，可诱导WRKY1和3的mRNA水平瞬时且极快速增加。相比之下，WRKY2的mRNA水平则伴随瞬时下降。WRKY mRNA水平对特定信号分子的这些快速变化表明，WRKY1、2和3在从激发子感知到PR1基因激活的信号转导途径中起关键作用。
