Desveaux D, Després C, Joyeux A, Subramaniam R, Brisson N
Department of Biochemistry, Université de Montréal, Montréal, Québec, Canada H3C 3J7.
Plant Cell. 2000 Aug;12(8):1477-89. doi: 10.1105/tpc.12.8.1477.
Elicitor-induced activation of the potato pathogenesis-related gene PR-10a requires a 30-bp promoter sequence termed the ERE (elicitor response element) that is bound by the nuclear factor PBF-2 (PR-10a binding factor 2). In this study, PBF-2 has been purified to near homogeneity from elicited tubers through a combination of anion-exchange and DNA affinity chromatography. Evidence demonstrates that inactive PBF-2 is stored in the nuclei of fresh tubers and becomes available for binding to the ERE upon elicitation. A protein with an apparent molecular mass of 24 kD (p24) is a DNA binding component of PBF-2. A cDNA encoding p24 has been cloned and encodes a novel protein with a potential transcriptional activation domain that could also act as a single-stranded DNA binding domain. Both PBF-2 and the cDNA-encoded protein bind with high affinity to the single-stranded form of the ERE in a sequence-specific manner. The inverted repeat sequence of the ERE, TGACAnnnnTGTCA, is critical for binding of this factor in vitro and for PR-10a expression in vivo, supporting the role of PBF-2 as a transcriptional regulator.
激发子诱导的马铃薯病程相关基因PR-10a的激活需要一个30 bp的启动子序列,即激发子反应元件(ERE),它可被核因子PBF-2(PR-10a结合因子2)结合。在本研究中,通过阴离子交换和DNA亲和层析相结合的方法,已从受激发的块茎中纯化出近乎纯一的PBF-2。有证据表明,无活性的PBF-2存储在新鲜块茎的细胞核中,激发后可与ERE结合。一种表观分子量为24 kD的蛋白质(p24)是PBF-2的DNA结合成分。已克隆出编码p24的cDNA,其编码一种具有潜在转录激活结构域的新型蛋白质,该结构域也可作为单链DNA结合结构域。PBF-2和cDNA编码的蛋白质均以序列特异性方式与ERE的单链形式高亲和力结合。ERE的反向重复序列TGACAnnnnTGTCA对于该因子在体外的结合以及体内PR-10a的表达至关重要,这支持了PBF-2作为转录调节因子的作用。