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水稻SUV3是一种能与DNA和RNA结合的双向解旋酶。

Rice SUV3 is a bidirectional helicase that binds both DNA and RNA.

作者信息

Tuteja Narendra, Tarique Mohammed, Tuteja Renu

出版信息

BMC Plant Biol. 2014 Oct 14;14:283. doi: 10.1186/s12870-014-0283-6.

Abstract

BACKGROUND

Helicases play crucial role in almost all the nucleic acid metabolism including replication, repair, recombination, transcription, translation, ribosome biogenesis and splicing and these processes regulate plant growth and development. It is suggested that helicases play essential roles in stabilizing growth in plants under stress because their presence in the stress-induced ORFs has been identified. Moreover in a recent study we have reported that SUV3 helicase from Oryza sativa (OsSUV3) functions in salinity stress tolerance in transgenic rice by improving the antioxidant machinery. SUV3 helicase has been identified and characterized from yeast and human systems but the properties and functions of plant SUV3 are poorly understood.

RESULTS

In this study, the purification and extensive characterization of recombinant OsSUV3 protein (67 kDa) is presented. OsSUV3 binds to DNA and RNA and exhibits DNA as well as RNA-dependent ATPase activities. It also contains the characteristic DNA and RNA helicase activity. OsSUV3 can use mainly ATP or dATP as energy source for the unwinding activity and it cannot unwind the blunt-end duplex DNA substrate. It is interesting to note that OsSUV3 unwinds DNA in both the 5'-3' and 3'-5 directions and thus its activity is bipolar in vitro. The Km values of OsSUV3 are 0.51 nM and 0.95 nM for DNA helicase and RNA helicase, respectively.

CONCLUSIONS

This study is the first direct evidence to show the bipolar DNA helicase activity of OsSUV3 protein. The unique properties of OsSUV3 including its dual helicase activity imply that it could be a multifunctional protein involved in biologically significant process of DNA and RNA metabolisms. These results should make significant contribution towards better understanding of SUV3 protein in plants.

摘要

背景

解旋酶在几乎所有核酸代谢过程中都发挥着关键作用,这些过程包括复制、修复、重组、转录、翻译、核糖体生物合成和剪接,并且这些过程调控着植物的生长发育。由于在胁迫诱导的开放阅读框中发现了解旋酶的存在,因此推测解旋酶在植物胁迫条件下稳定生长中发挥着重要作用。此外,在最近的一项研究中我们报道,来自水稻的SUV3解旋酶(OsSUV3)通过改善抗氧化机制在转基因水稻的耐盐胁迫中发挥作用。SUV3解旋酶已在酵母和人类系统中得到鉴定和表征,但对植物SUV3的特性和功能了解甚少。

结果

在本研究中,展示了重组OsSUV3蛋白(67 kDa)的纯化及广泛表征。OsSUV3与DNA和RNA结合,并表现出DNA以及RNA依赖性ATP酶活性。它还具有特征性的DNA和RNA解旋酶活性。OsSUV3主要可以使用ATP或dATP作为解旋活性的能量来源,并且它不能解开平端双链DNA底物。有趣的是,OsSUV3在5'-3'和3'-5'两个方向上都能解开DNA,因此其活性在体外是双向的。OsSUV3对DNA解旋酶和RNA解旋酶的Km值分别为0.51 nM和0.95 nM。

结论

本研究是首个直接证据,证明了OsSUV3蛋白具有双向DNA解旋酶活性。OsSUV3的独特特性,包括其双重解旋酶活性,意味着它可能是一种参与DNA和RNA代谢重要生物学过程的多功能蛋白。这些结果将为更好地理解植物中的SUV3蛋白做出重要贡献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2dfa/4207899/638b7edb38a2/12870_2014_283_Fig1_HTML.jpg

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