Structural Basis for Substrate Recognition by the Editing Domain of Isoleucyl-tRNA Synthetase

Ryuya Fukunaga, Shigeyuki Yokoyama

Research output: Contribution to journalArticle

Abstract

In isoleucyl-tRNA synthetase (IleRS), the "editing" domain contributes to accurate aminoacylation by hydrolyzing the mis-synthesized intermediate, valyl-adenylate, in the "pre-transfer" editing mode and the incorrect final product, valyl-tRNAIle, in the "post-transfer" editing mode. In the present study, we determined the crystal structures of the Thermus thermophilus IleRS editing domain complexed with the substrate analogues in the pre and post-transfer modes, both at 1.7 Å resolution. The active site accommodates the two analogues differently, with the valine side-chain rotated by about 120° and the adenosine moiety oriented upside down. The substrate-binding pocket adjusts to the adenosine-monophosphate and adenosine moieties in the pre and post-transfer modes, respectively, by flipping the Trp227 side-chain by about 180°. The substrate recognition mechanisms of IleRS are characterized by the active-site rearrangement between the two editing modes, and therefore differ from those of the homologous valyl and leucyl-tRNA synthetases from T. thermophilus, in which the post-transfer mode is predominant. Both modes of editing activities were reduced by replacements of Trp227 with Ala, Val, Leu, and His, but not by those with Phe and Tyr, indicating that the aromatic ring of Trp227 is important for the substrate recognition. In both editing modes, Thr233 and His319 recognize the substrate valine side-chain, regardless of the valine side-chain rotation, and reject the isoleucine side-chain. The T233A and H319A mutants have detectable editing activities against the cognate isoleucine.

Original languageEnglish (US)
Pages (from-to)901-912
Number of pages12
JournalJournal of Molecular Biology
Volume359
Issue number4
DOIs
StatePublished - Jun 16 2006
Externally publishedYes

Fingerprint

Isoleucine-tRNA Ligase
Valine
Thermus thermophilus
Isoleucine
Catalytic Domain
Valine-tRNA Ligase
RNA, Transfer, Ile
Aminoacylation
Adenosine

Keywords

  • amino acid
  • aminoacyl-tRNA synthetase
  • editing
  • isoleucine
  • valine

ASJC Scopus subject areas

  • Virology

Cite this

Structural Basis for Substrate Recognition by the Editing Domain of Isoleucyl-tRNA Synthetase. / Fukunaga, Ryuya; Yokoyama, Shigeyuki.

In: Journal of Molecular Biology, Vol. 359, No. 4, 16.06.2006, p. 901-912.

Research output: Contribution to journalArticle

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