Breaking the stereo barrier of amino acid attachment to tRNA by a single nucleotide.
Aminoacyl-tRNA synthetases are responsible for attaching amino acid residues to the tRNA 3'-end. The two classes of synthetases approach tRNA as mirror images, with opposite but symmetrical stereochemistries that allow the class I enzymes to attach amino acid residues to the 2'-hydroxyl group of the terminal ribose, whereas, the class II enzymes attach amino acid residues to the 3'-hydroxyl group. However, we show here that the attachment of cysteine to tRNA(Cys) by the class I cysteinyl-tRNA synthetase (CysRS) is flexible; the enzyme is capable of using either the 2' or 3'-hydroxyl group as the attachment site. The molecular basis for this flexibility was investigated. Introduction of the nucleotide U73 of tRNA(Cys) into tRNA(Val) was found to confer the flexibility. While valylation of the wild-type tRNA(Val) by the class I ValRS was strictly dependent on the terminal 2'-hydroxyl group, that of the U73 mutant of tRNA(Val) occurred at either the 2' or 3'-hydroxyl group. Thus, the single nucleotide U73 of tRNA has the ability to break the stereo barrier of amino acid attachment to tRNA, by mobilizing the 2' and 3'-hydroxyl groups of A76 in flexible geometry with respect to the tRNA acceptor stem.
Shitivelband
Svetlana
S
Department of Biochemistry and Molecular Pharmacology, Thomas Jefferson University, 233 South 10th Street, Philadelphia, PA 19107, USA.
Hou
Ya-Ming
YM
D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism*; D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000737:chemistry / Q000378:metabolism*; D012350:RNA, Transfer, Cys / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012366:RNA, Transfer, Val / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D000596:Amino Acids; D029968:Escherichia coli Proteins; D009711:Nucleotides; D012350:RNA, Transfer, Cys; D012366:RNA, Transfer, Val; D000604:Amino Acyl-tRNA Synthetases; C019975:cysteinyl-tRNA synthetase