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大麦Nudix水解酶基因的结构与分子特征

Structure and molecular characterization of barley nudix hydrolase genes.

作者信息

Tanaka Sayuri, Kihara Makoto, Sugimoto Manabu

机构信息

a Institute of Plant Science and Resources , Okayama University , Okayama , Japan.

出版信息

Biosci Biotechnol Biochem. 2015;79(3):394-401. doi: 10.1080/09168451.2014.978259. Epub 2014 Nov 7.

Abstract

Putative nudix hydrolase (NUDX) genes, which encode amino acid sequences showing homology with those of Arabidopsis NUDXs and conserve nudix motif, were identified from barley. The 14 deduced barley NUDXs (HvNUDX1-14) were classified into established subfamilies, except for 8-oxo-deoxyguanosine 5'-triphosphate (8-oxo-dGTP) pyrophosphohydrolase and mRNA decapping enzyme subfamilies, and three substrate-unknown subfamilies. Drought and UV-C stresses, respectively, up-regulated 7 and 4 HvNUDX genes, but some homologs of Arabidopsis NUDXs showed different responses to abiotic stress. HvNUDX12 gene, belonging to diadenosine tetraphosphates (Ap₄A) pyrophosphohydrolase subfamily gene and up-regulated by UV-C, was expressed in Escherichia coli cells. The recombinant protein showed 8-oxo-dGTP, Ap₄A, and guanosine-3',5'-tetraphosphate (ppGpp) pyrophosphohydrolase activities, and the suppression of the lacZ amber mutation in a mutT-deficient E. coli cells caused by the incorporation of 8-oxo-GTP into mRNA was prevented to a significant degree. These results suggest that barley NUDXs have unique constitution and response of NUDX to abiotic stress.

摘要

从大麦中鉴定出了假定的Nudix水解酶(NUDX)基因,这些基因编码的氨基酸序列与拟南芥NUDX的氨基酸序列具有同源性,并保守了Nudix基序。除8-氧代脱氧鸟苷5'-三磷酸(8-氧代-dGTP)焦磷酸水解酶和mRNA去帽酶亚家族以及三个底物未知的亚家族外,14个推导的大麦NUDX(HvNUDX1-14)被分类到已确立的亚家族中。干旱和UV-C胁迫分别上调了7个和4个HvNUDX基因,但拟南芥NUDX的一些同源物对非生物胁迫表现出不同的反应。属于二腺苷四磷酸(Ap₄A)焦磷酸水解酶亚家族基因且被UV-C上调的HvNUDX12基因在大肠杆菌细胞中表达。重组蛋白表现出8-氧代-dGTP、Ap₄A和鸟苷-3',5'-四磷酸(ppGpp)焦磷酸水解酶活性,并且在很大程度上阻止了由于8-氧代-GTP掺入mRNA而导致的mutT缺陷型大肠杆菌细胞中lacZ琥珀突变的抑制。这些结果表明大麦NUDX具有独特的组成以及NUDX对非生物胁迫的反应。

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