Golovina Anna Y, Sergiev Petr V, Golovin Andrey V, Serebryakova Marina V, Demina Irina, Govorun Vadim M, Dontsova Olga A
Department of Chemistry, Moscow State University, Moscow 119992, Russia.
RNA. 2009 Jun;15(6):1134-41. doi: 10.1261/rna.1494409. Epub 2009 Apr 21.
Transfer RNA is highly modified. Nucleotide 37 of the anticodon loop is represented by various modified nucleotides. In Escherichia coli, the valine-specific tRNA (cmo(5)UAC) contains a unique modification, N(6)-methyladenosine, at position 37; however, the enzyme responsible for this modification is unknown. Here we demonstrate that the yfiC gene of E. coli encodes an enzyme responsible for the methylation of A37 in tRNA(1)(Val). Inactivation of yfiC gene abolishes m(6)A formation in tRNA(1)(Val), while expression of the yfiC gene from a plasmid restores the modification. Additionally, unmodified tRNA(1)(Val) can be methylated by recombinant YfiC protein in vitro. Although the methylation of m(6)A in tRNA(1)(Val) by YfiC has little influence on the cell growth under standard conditions, the yfiC gene confers a growth advantage under conditions of osmotic and oxidative stress.
转运RNA经过高度修饰。反密码子环的第37位核苷酸由多种修饰核苷酸代表。在大肠杆菌中,缬氨酸特异性tRNA(cmo(5)UAC)在第37位含有一种独特的修饰,即N(6)-甲基腺苷;然而,负责这种修饰的酶尚不清楚。在这里,我们证明大肠杆菌的yfiC基因编码一种负责tRNA(1)(Val)中A37甲基化的酶。yfiC基因的失活消除了tRNA(1)(Val)中m(6)A的形成,而从质粒表达yfiC基因可恢复这种修饰。此外,未修饰的tRNA(1)(Val)在体外可被重组YfiC蛋白甲基化。虽然在标准条件下YfiC对tRNA(1)(Val)中m(6)A的甲基化对细胞生长影响不大,但yfiC基因在渗透和氧化应激条件下赋予生长优势。