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葡萄(Vitis vinifera)S-腺苷甲硫氨酸脱羧酶的克隆、功能鉴定及结构建模

Cloning, functional identification and structural modelling of Vitis vinifera S-adenosylmethionine decarboxylase.

作者信息

Tassoni Annalisa, Franceschetti Marina, Tasco Gianluca, Casadio Rita, Bagni Nello

机构信息

Department of Biology e s and Interdepartmental Centre for Biotechnology, University of Bologna, Via Irnerio 42, Bologna, Italy.

出版信息

J Plant Physiol. 2007 Sep;164(9):1208-19. doi: 10.1016/j.jplph.2006.07.009. Epub 2006 Sep 18.

Abstract

In this paper we report the cloning and full sequencing of S-adenosylmethionine decarboxylase (SAMDC, EC 4.1.1.50) cDNA from Vitis vinifera L. (VV) leaves, an enzyme belonging to the polyamine biosynthetic pathway, which appears to play an important role in the regulation of plant growth and development. The presence of two overlapping ORFs (tiny ORF and small ORF) upstream of the main ORF is reported in the Vitis cDNA. When the Vitis SAMDC cDNA was expressed in yeast without the two upstream ORFs, the resulting activity was about 50 times higher than the activity obtained with the full cDNA. These results demonstrated the strong regulatory activity of the tiny and small ORFs. RT-PCR expression analysis showed evidence of a similar mRNA level in all the tissues tested, with the exception of the petioles. The VV SAMDC was also modelled using its homologues from Solanum tuberosum and Homo sapiens as template. The present work confirmed, for the first time in a woody plant of worldwide economic interest such as grapevine, the presence of a regulatory mechanism of SAMDC, enzyme that has a well-established importance in the modulation of plant growth and development.

摘要

在本文中,我们报道了从葡萄(Vitis vinifera L.,VV)叶片中克隆并对S-腺苷甲硫氨酸脱羧酶(SAMDC,EC 4.1.1.50)cDNA进行全序列分析,该酶属于多胺生物合成途径,似乎在植物生长和发育的调控中发挥重要作用。葡萄cDNA中报道了在主要开放阅读框(ORF)上游存在两个重叠的ORF(微小ORF和小ORF)。当葡萄SAMDC cDNA在没有两个上游ORF的情况下在酵母中表达时,产生的活性比用完整cDNA获得的活性高约50倍。这些结果证明了微小和小ORF具有强大的调控活性。逆转录聚合酶链反应(RT-PCR)表达分析表明,除叶柄外,在所有测试组织中mRNA水平相似。还以马铃薯和人类的同源物为模板对VV SAMDC进行了建模。本研究首次在具有全球经济价值的木本植物葡萄中证实了SAMDC的调控机制的存在,该酶在植物生长和发育的调节中具有公认的重要性。

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