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三个桃脱水素基因的表达比较及转录起始

Comparative expression and transcript initiation of three peach dehydrin genes.

作者信息

Bassett Carole Leavel, Wisniewski Michael E, Artlip Timothy S, Richart Greg, Norelli John L, Farrell Robert E

机构信息

USDA, ARS, Appalachian Fruit Research Station, 2217 Wiltshire Road, Kearneysville, WV 25430, USA.

出版信息

Planta. 2009 Jun;230(1):107-18. doi: 10.1007/s00425-009-0927-1. Epub 2009 Apr 10.

Abstract

Dehydrin genes encode proteins with demonstrated cryoprotective and antifreeze activity, and they respond to a variety of abiotic stress conditions that have dehydration as a common component. Two dehydrins from peach (Prunus persica L. [Batsch.]) have been previously characterized; here, we describe the characterization of a third dehydrin from peach bark, PpDhn3, isolated by its response to low temperature. The expression of all three dehydrin genes was profiled by semi-quantitative reverse transcription PCR, and transcript initiation was mapped for all three genes using the RNA ligase-mediated 5' rapid amplification of cDNA ends technique. PpDhn3 transcripts from bark collected in December or July, as well as transcripts from developing fruit, initiated at a single site. Although most of the PpDhn1 transcripts initiated at a similar position, those from young fruit initiated much further upstream of the consensus TATA box. Bark and fruit transcripts encoding PpDhn2 initiated ca. 30 bases downstream of a consensus TATA box; however, transcripts from ripe fruit initiated further upstream. Ripe fruit transcripts of PpDhn2 contain a 5' leader intron which is predicted to add some 34 amino acids to the N-terminal methionine of the cognate protein when properly processed. Secondary structure prediction of sequences surrounding the TATA box suggests that conformational transitions associated with decreasing temperature contribute to the regulation of expression of the cold-responsive dehydrin genes. Taken together these results reveal new, unexpected levels of gene regulation contributing to the overall expression pattern of peach dehydrins.

摘要

脱水素基因编码的蛋白质具有已证实的冷冻保护和抗冻活性,它们对多种以脱水为共同成分的非生物胁迫条件作出反应。此前已对桃(Prunus persica L. [Batsch.])中的两种脱水素进行了表征;在此,我们描述了从桃树皮中分离出的第三种脱水素PpDhn3的表征,它是通过对低温的反应而分离得到的。通过半定量逆转录PCR分析了所有三种脱水素基因的表达,并使用RNA连接酶介导的cDNA末端5'快速扩增技术对所有三个基因的转录起始位点进行了定位。12月或7月采集的树皮中的PpDhn3转录本,以及发育中的果实中的转录本,均起始于单个位点。虽然大多数PpDhn1转录本起始于相似位置,但幼果中的转录本起始于共有TATA框上游更远的位置。编码PpDhn2的树皮和果实转录本起始于共有TATA框下游约30个碱基处;然而,成熟果实中的转录本起始于更上游。PpDhn2的成熟果实转录本包含一个5'前导内含子,预计在正确加工时会在同源蛋白的N端甲硫氨酸上添加约34个氨基酸。TATA框周围序列的二级结构预测表明,与温度降低相关的构象转变有助于调控冷响应脱水素基因的表达。综合这些结果揭示了新的、意想不到的基因调控水平,这些调控水平有助于桃脱水素的整体表达模式。

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