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用于控制光介导表达的胡黄连3-羟基-3-甲基戊二酰辅酶A还原酶启动子中的GATA和SORLIP基序。

The GATA and SORLIP motifs in the 3-hydroxy-3-methylglutaryl-CoA reductase promoter of Picrorhiza kurrooa for the control of light-mediated expression.

作者信息

Kawoosa Tabasum, Gahlan Parul, Devi Aribam Surbala, Kumar Sanjay

机构信息

Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology (Council of Scientific and Industrial Research), P.O. Box No. 6, Palampur, 176061, Himachal Pradesh, India,

出版信息

Funct Integr Genomics. 2014 Mar;14(1):191-203. doi: 10.1007/s10142-013-0350-3. Epub 2013 Dec 7.

Abstract

Light upregulates the expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) in Picrorhiza kurrooa, an endangered medicinal herb. Upstream sequences of HMGR of P. kurrooa (PropkHMGR) were analyzed in relation to its role in light-mediated regulation of gene expression. GATA motif in PropkHMGR exhibited stronger DNA-protein interaction with the nuclear extract of dark-exposed plants in contrast to SORLIP that exhibited stronger binding with the nuclear extract of light-exposed plants. Analysis of PropkHMGR (PropkHMGR-D1, -1,059/-1) and its deletion fragments PropkHMGR-D2 (-825/-1), PropkHMGR-D3 (-651/-1), PropkHMGR-D4 (-452/-1), and PropkHMGR-D5 (-101/-1) in Arabidopsis thaliana showed PropkHMGR to regulate gene expression [β-glucuronidase (GUS) was used as a reporter gene] at all the developmental stages but only in actively dividing tissues, excluding anthers. Whereas, PropkHMGR-D2 regulated GUS expression in relatively older seedlings but the expression was observed only in shoot apical meristem, root tips, and anthers. PropkHMGR-mediated gene expression was higher in dark as compared to that in the light in Arabidopsis across four temperatures studied. As opposed to the results in P. kurrooa, GATA motifs exhibited DNA-protein interaction with nuclear extract of light-exposed plants of Arabidopsis. SORLIP motifs in Arabidopsis also exhibited DNA-protein interaction with nuclear extract of light-exposed plants as in P. kurrooa. Data showed that (1) PropkHMGR regulated light-mediated gene expression and (2) GATA motif exhibited an inverse relationship between strength of DNA-protein interaction and the gene expression whereas the relationship was species specific for SORLIP.

摘要

光照可上调濒危药用植物胡黄连中3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)的表达。对胡黄连HMGR(PropkHMGR)的上游序列进行了分析,以研究其在光照介导的基因表达调控中的作用。与SORLIP相比,PropkHMGR中的GATA基序与黑暗处理植物的核提取物表现出更强的DNA-蛋白质相互作用,而SORLIP与光照处理植物的核提取物结合更强。对拟南芥中的PropkHMGR(PropkHMGR-D1,-1,059/-1)及其缺失片段PropkHMGR-D2(-825/-1)、PropkHMGR-D3(-651/-1)、PropkHMGR-D4(-452/-1)和PropkHMGR-D5(-101/-1)进行分析,结果表明PropkHMGR在所有发育阶段均能调控基因表达[以β-葡萄糖醛酸酶(GUS)作为报告基因],但仅在活跃分裂的组织中,花药除外。然而,PropkHMGR-D2在相对较老的幼苗中调控GUS表达,但仅在茎尖分生组织、根尖和花药中观察到该表达。在研究的四个温度下,与光照条件下相比,PropkHMGR介导的基因表达在黑暗条件下的拟南芥中更高。与胡黄连的结果相反,GATA基序与光照处理的拟南芥植物的核提取物表现出DNA-蛋白质相互作用。拟南芥中的SORLIP基序也与光照处理的植物的核提取物表现出DNA-蛋白质相互作用,如同在胡黄连中一样。数据表明:(1)PropkHMGR调控光照介导的基因表达;(2)GATA基序在DNA-蛋白质相互作用强度与基因表达之间呈现反比关系,而SORLIP的这种关系具有物种特异性。

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