Richard S, Drevet C, Jouanin L, Séguin A
Centre de recherche en biologie forestière, Pavillon C.-E.-Marchand, Université Laval, Sainte-Foy, Canada.
Gene. 1999 Nov 29;240(2):379-88. doi: 10.1016/s0378-1119(99)00426-6.
A presumably full-length cDNA encoding a putative glycine-rich RNA binding protein was isolated from a lambdaZAP cDNA library prepared from mRNAs extracted from needles of 2year old white spruce seedlings, which had been either wounded or jasmonate-treated. The cDNA, designated PgRNP (Picea glauca RNP protein), presents a 468bp open reading frame encoding a 155 amino acid protein. This polypeptide possesses an RNA binding domain (RNP-CS) and a glycine-rich domain. Comparative alignment reveals extensive homologies to glycine-rich RNA binding proteins containing an RNP-CS found in other angiosperm species. Genomic hybridization experiments suggest that the PgRNP gene is part of a small multigene family with at least four members. RNA blot analysis revealed that the PgRNP transcript is expressed in all tissues from non-stressed plants. Constitutive mRNA level was found in needle tissue from control as well as methyl-jasmonate treated plants. Wounding had no clear induction effect. Jasmonic acid treatment and systemic wound response had a positive effect on transcript accumulation. Transcript accumulation was slightly induced by cold in needles, and repressed by drought stress in both needle and root tissues of 2year old plants. Finally, the level of PgRNP accumulation was induced by wounding and repressed in 2week old dark-grown seedlings upon jasmonate treatments.
从一个λZAP cDNA文库中分离出一个推测为全长的cDNA,该文库是用从2年生白云杉幼苗针叶中提取的mRNA构建的,这些幼苗要么受过伤,要么经过茉莉酸处理。该cDNA命名为PgRNP(白云杉RNP蛋白),具有一个468bp的开放阅读框,编码一个155个氨基酸的蛋白质。该多肽具有一个RNA结合结构域(RNP-CS)和一个富含甘氨酸的结构域。比较比对显示,它与其他被子植物物种中发现的含有RNP-CS的富含甘氨酸的RNA结合蛋白有广泛的同源性。基因组杂交实验表明,PgRNP基因是一个至少有四个成员的小多基因家族的一部分。RNA印迹分析显示,PgRNP转录本在未受胁迫植物的所有组织中均有表达。在对照植物以及经茉莉酸甲酯处理的植物的针叶组织中发现了组成型mRNA水平。创伤没有明显的诱导作用。茉莉酸处理和系统性创伤反应对转录本积累有积极影响。在针叶中,低温轻微诱导转录本积累,而在2年生植物的针叶和根组织中,干旱胁迫则抑制转录本积累。最后,在2周龄黑暗生长的幼苗中,创伤诱导了PgRNP的积累水平,而茉莉酸处理则抑制了其积累。