Bilokapic Silvija, Korencic Dragana, Söll Dieter, Weygand-Durasevic Ivana
Department of Chemistry, Faculty of Science, University of Zagreb, Croatia.
Eur J Biochem. 2004 Feb;271(4):694-702. doi: 10.1111/j.1432-1033.2003.03971.x.
The methanogenic archaea Methanococcus jannaschii and M. maripaludis contain an atypical seryl-tRNA synthetase (SerRS), which recognizes eukaryotic and bacterial tRNAsSer, in addition to the homologous tRNASer and tRNASec species. The relative flexibility in tRNA recognition displayed by methanogenic SerRSs, shown by aminoacylation and gel mobility shift assays, indicates the conservation of some serine determinants in all three domains. The complex of M. maripaludis SerRS with the homologues tRNASer was isolated by gel filtration chromatography. Complex formation strongly depends on the conformation of tRNA. Therefore, the renaturation conditions for in vitro transcribed tRNASer(GCU) isoacceptor were studied carefully. This tRNA, unlike many other tRNAs, is prone to dimerization, possibly due to several stretches of complementary oligonucleotides within its sequence. Dimerization is facilitated by increased tRNA concentration and can be diminished by fast renaturation in the presence of 5 mm magnesium chloride.
产甲烷古菌詹氏甲烷球菌(Methanococcus jannaschii)和泥沼甲烷球菌(M. maripaludis)含有一种非典型的丝氨酸 - tRNA合成酶(SerRS),除了同源的tRNASer和tRNASec种类外,它还能识别真核生物和细菌的tRNAsSer。通过氨酰化和凝胶迁移率变动分析表明,产甲烷SerRSs在tRNA识别方面表现出相对灵活性,这表明在所有三个结构域中某些丝氨酸决定簇是保守的。通过凝胶过滤色谱法分离出泥沼甲烷球菌SerRS与同源tRNASer的复合物。复合物的形成强烈依赖于tRNA的构象。因此,对体外转录的tRNASer(GCU)同功受体的复性条件进行了仔细研究。与许多其他tRNA不同,这种tRNA容易发生二聚化,这可能是由于其序列中存在几段互补寡核苷酸。tRNA浓度增加会促进二聚化,而在5 mM氯化镁存在下快速复性可减少二聚化。