EVIDENCE FOR RNA-OLIGONUCLEOTIDES IN PLANT VACUOLES ISOLATED FROM CULTURED TOMATO CELLS
EVIDENCE FOR RNA-OLIGONUCLEOTIDES IN PLANT VACUOLES ISOLATED FROM CULTURED TOMATO CELLS
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DOI：
10.1104/pp.94.3.1163
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发表时间：
1990-11-01
期刊：
影响因子：
7.4
通讯作者：
GLUND, K
中科院分区：
文献类型：
--
作者：
ABEL, S;BLUME, B;GLUND, K
我们已经表明，高度纯化的悬浮培养番茄液泡，（Lycopersicon esculentum）细胞含有RNA-寡核苷酸，使用两种不同的方法来标记和检测RNA：（a）用[5- 3 H]尿苷活体标记细胞RNA，随后从原生质体制备液泡并定量放射性标记的物质;和（B）从番茄液泡制备的核酸的体外标记和测序凝胶上的分析以及它们作为RNA的鉴定。在液泡制剂中发现的RNA的液泡内位置的结论是从分析RNA完整的细胞器后，重复的浮选步骤，以及核糖核酸酶A处理。原生质体中约3%的RNA定位于液泡内，超过未裂解的原生质体和亚细胞器污染所作的贡献的几倍。空泡RNA的大小分布的调查显示，寡核苷酸模式显着不同，这将产生污染的原生质体，空泡RNA片段大大短于80个核苷酸。这些寡核糖核苷酸（3′-磷酸化末端;相对富含嘧啶）可能是番茄液泡核糖核酸酶I作用的产物，此外，在裂解的液泡制备物中，液泡RNA通过固有酶活性的酶水解支持植物液泡参与细胞核溶解过程的假设。
We have shown that highly purified vacuoles of suspension-cultured tomato (Lycopersicon esculentum) cells contain RNA-oligonucleotides, using two different approaches to label and detect RNA: (a)in vivolabeling of cellular RNA with [5-3H]uridine, followed by preparation of vacuoles from protoplasts and by quantification of radioactively labeled material; and (b)in vitrolabeling and analysis on sequencing gels of nucleic acids prepared from tomato vacuoles and their identification as RNA. The intravacuolar location of the RNA found in vacuolar preparations was concluded from analyzing for RNA intact organelles after repeated flotation steps as well as ribonuclease A treatment. About 3% of the RNA in protoplasts was localized within vacuoles, exceeding by severalfold the contribution made by contamination with unlysed protoplasts and subcellular organelles. Investigation of the size distribution of vacuolar RNA revealed an oligonucleotide pattern strikingly different from that which would arise from contaminating protoplasts; vacuolar RNA fragments are considerably shorter than 80 nucleotides. Characterization of these oligoribonucleotides (3′-phosphorylated termini; relatively rich in pyrimidines) as possible products of tomato vacuolar ribonuclease I action, and, in addition, enzymatic hydrolysis of vacuolar RNA by inherent enzyme activities in lysed vacuole preparations support the hypothesis that plant vacuoles are involved in cellular nucleolytic processes.