Nishizawa Y, Kawakami A, Hibi T, He D Y, Shibuya N, Minami E
Department of Biotechnology, National Institute of Agrobiological Resources, Tsukuba, Japan.
Plant Mol Biol. 1999 Mar;39(5):907-14. doi: 10.1023/a:1006161802334.
Expression patterns of chitinase transcripts induced by N-acetylchitooligosaccharide elicitor were analyzed by northern blot hybridization in order to reveal a signal transduction pathway leading to the activation of class I chitinase genes (Cht-1 and Cht-3), which may play an important role in producing N-acetylchitooligosaccharide elicitor. The transcription level of both genes was enhanced in response to N-acetylchitooligosaccharides larger than pentaose at subnanomolar concentrations. These structure and dose dependencies were consistent not only with those for a 75 kDa high-affinity binding protein for N-acetylchitooligosaccharide elicitor in the plasma membrane, but also with other series of cellular responses including phytoalexin production and the expression of elicitor-responsive genes (EL2, EL3). Therefore, the elicitor signal to evoke these cellular responses including the activation of the chitinase genes could be common and transmitted into cells through the 75 kDa protein. However, the signal transduction pathway for the activation of the chitinase gene appeared to diverge from those for the other elicitor-responsive genes shortly after the signal perception. It was shown that the induction of chitinase expression by N-acetylchitooligosaccharide would require protein phosphorylation, but not de novo protein synthesis. The oxidative burst was demonstrated not to be necessary for transcriptional induction of the all four elicitor-responsive genes (Cht, PAL, EL2, EL3) by N-acetylchitooligosaccharide.
为了揭示导致I类几丁质酶基因(Cht-1和Cht-3)激活的信号转导途径,通过Northern印迹杂交分析了由N-乙酰壳寡糖激发子诱导的几丁质酶转录本的表达模式,这可能在产生N-乙酰壳寡糖激发子中起重要作用。在亚纳摩尔浓度下,大于五糖的N-乙酰壳寡糖可增强这两个基因的转录水平。这些结构和剂量依赖性不仅与质膜中N-乙酰壳寡糖激发子的75 kDa高亲和力结合蛋白的结构和剂量依赖性一致,而且与包括植保素产生和激发子响应基因(EL2、EL3)表达在内的其他一系列细胞反应一致。因此,引发这些细胞反应(包括几丁质酶基因激活)的激发子信号可能是共同的,并通过75 kDa蛋白传递到细胞中。然而,在信号感知后不久,几丁质酶基因激活的信号转导途径似乎与其他激发子响应基因的信号转导途径不同。结果表明,N-乙酰壳寡糖诱导几丁质酶表达需要蛋白质磷酸化,但不需要从头合成蛋白质。已证明氧化爆发对于N-乙酰壳寡糖对所有四个激发子响应基因(Cht、PAL、EL2、EL3)的转录诱导不是必需的。