Characterization of Arabidopsis thaliana cDNAs that render yeasts tolerant toward the thiol-oxidizing drug diamide.
Characterization of Arabidopsis thaliana cDNAs that render yeasts tolerant toward the thiol-oxidizing drug diamide.
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拟南芥 cDNA 的表征，使酵母对硫醇氧化药物二酰胺具有耐受性。
DOI：
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发表时间：
1995
影响因子：
11.1
通讯作者：
Dirk Inzé
中科院分区：
文献类型：
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作者：
S. Kushnir;E. Babiychuk;K. Kampfenkel;E. Belles;M. V. Montagu;Dirk Inzé
二胺氧化细胞硫醇并诱导氧化应激。为了分离出过表达可增加植物抗氧化损伤能力的基因，对酵母菌进行了体内二胺耐受性筛选。利用酵母表达载体中的拟南芥cDNA文库转化了一株具有完整抗氧化防御的酵母菌株。从大约10(5)个初级转化子中选择细胞对二胺具有抗性。分离了三个具有二胺耐受性的拟南芥cdna。这种药物耐受性是特异性的，对氢过氧化物没有交叉耐受性。一个cDNA （D3）编码一个多肽，该多肽具有不同DnaJ伴侣家族的氨基末端J结构域特征。另一个（D18）编码假定的dtdp -d -葡萄糖4,6-脱水酶。令人惊讶的是，第三个cDNA （D22）编码了γ -谷氨酰转移酶的植物同源物。在细胞硫醇耗尽的条件下，很难预测这些基因的表达会导致存活率的提高。因此，我们认为这种克隆方法可能有助于分离出有助于应对氧化应激的植物基因。
Diamide oxidizes cellular thiols and induces oxidative stress. To isolate plant genes which may, when overexpressed, increase tolerance of plants toward oxidative damage, an in vivo diamide tolerance screening in yeasts was used. An Arabidopsis cDNA library in a yeast expression vector was used to transform a yeast strain with intact antioxidant defense. Cells from approximately 10(5) primary transformants were selected for resistance to diamide. Three Arabidopsis cDNAs which confer diamide tolerance were isolated. This drug tolerance was specific and no cross tolerance toward hydroperoxides was found. One cDNA (D3) encodes a polypeptide which has an amino-terminal J domain characteristic of a divergent family of DnaJ chaperones. Another (D18) encodes a putative dTDP-D-glucose 4,6-dehydratase. Surprisingly, the third cDNA (D22) encodes a plant homolog of gamma-glutamyltransferases. It would have been difficult to predict that the expression of those genes would lead to an improved survival under conditions of depletion of cellular thiols. Hence, we suggest that this cloning approach may be a useful contribution to the isolation of plant genes that can help to cope with oxidative stress.