Takane K, Tajima S, Kouchi H
Department of Agriculture, Kagawa University, Japan.
Mol Plant Microbe Interact. 1997 Aug;10(6):735-41. doi: 10.1094/MPMI.1997.10.6.735.
Nodule-specific uricase (uricase II) is a homotetramer of a 33-kDa polypeptide, nodulin 35, and plays a key role in the assimilation of nitrogen fixed by microsymbionts in most legumes that have determinate nodules. We have isolated two distinct genes, UR2 and UR9, that encode for nodulin 35 from a soybean genomic library. Their corresponding cDNAs were also isolated from a nodule cDNA library. UR2 and UR9 both encode for 309 amino acid proteins with 12 amino acid differences. The expression of these two genes in various organs of soybean was examined by reverse transcription-polymerase chain reaction with primers specific to each cDNA sequences. Expression of UR9 was almost specific in root nodules, although it was expressed in roots, primary leaves, and developing seed at very low levels. In contrast, the UR2 transcripts were present in almost all plant organs at low levels, but no enhancement of the expression was observed in nodules. Thus, UR9 behaves as a nodulin gene, whereas UR2 is a nonsymbiotic uricase II gene. The sequences of their potential promoter regions share high homology within regions up to about 400 bp upstream from the translation initiation sites. These results suggest that symbiotic and nonsymbiotic uricase II genes diverged by gene duplication and that relatively small alterations in the promoter sequence enable the nodule-specific expression.
根瘤特异性尿酸酶(尿酸酶II)是一种由33 kDa多肽结节蛋白35组成的同四聚体,在大多数具有有限根瘤的豆科植物中,它在微共生体固定氮的同化过程中起关键作用。我们从大豆基因组文库中分离出了两个不同的基因UR2和UR9,它们编码结节蛋白35。它们相应的cDNA也从根瘤cDNA文库中分离出来。UR2和UR9都编码309个氨基酸的蛋白质,两者有12个氨基酸的差异。用针对每个cDNA序列的特异性引物,通过逆转录-聚合酶链反应检测了这两个基因在大豆各器官中的表达。UR9的表达几乎只在根瘤中出现,尽管它在根、初生叶和发育中的种子中也有极低水平的表达。相比之下,UR2转录本在几乎所有植物器官中都有低水平表达,但在根瘤中未观察到表达增强。因此,UR9表现为一个根瘤蛋白基因,而UR2是一个非共生尿酸酶II基因。它们潜在启动子区域的序列在翻译起始位点上游约400 bp以内的区域具有高度同源性。这些结果表明,共生和非共生尿酸酶II基因通过基因复制而分化,并且启动子序列中相对较小的改变就能实现根瘤特异性表达。