Activation of the Pyrrolysine Suppressor tRNA Requires Formation of a Ternary Complex with Class I and Class II Lysyl-tRNA Synthetases

Authors

Carla Polycarpo, Yale University
Alexandre Ambrogelly, Yale University
Benfang Ruan, Yale University
Debra Tumbula-Hansen, Yale University
Sandro F. Ataide, The Ohio State University
Ryuichiro Ishitani, Tokyo Institute of Technology
Shigeyuki Yokoyama, Tokyo Institute of Technology
Osamu Nureki, Tokyo Institute of Technology
Michael Ibba, Chapman UniversityFollow
Dieter Söll, Yale University

Document Type

Article

Publication Date

8-28-2003

Abstract

Monomethylamine methyltransferase of the archaeon Methanosarcina barkeri contains a rare amino acid, pyrrolysine, encoded by the termination codon UAG. Translation of this UAG requires the aminoacylation of the corresponding amber suppressor tRNAPyl. Previous studies reported that tRNAPyl could be aminoacylated by the synthetase-like protein PylS. We now show that tRNAPyl is efficiently aminoacylated in the presence of both the class I LysRS and class II LysRS of M. barkeri, but not by either enzyme acting alone or by PylS. In vitro studies show that both the class I and II LysRS enzymes must bind tRNAPyl in order for the aminoacylation reaction to proceed. Structural modeling and selective inhibition experiments indicate that the class I and II LysRSs form a ternary complex with tRNAPyl, with the aminoacylation activity residing in the class II enzyme.

Comments

This article was originally published in Molecular Cell, volume 12, in 2003. https://doi.org/10.1016/S1097-2765(03)00280-6

Recommended Citation

Polycarpo, C., Ambrogelly, A., Ruan, B., Tumbula-Hansen, D., Ataide, S.F., Ishitani, R., Yokoyama, S., Nureki, O., Ibba, M. and Söll, D. (2003) Activation of the pyrrolysine suppressor tRNA requires formation of a ternary complex with class I and class II lysyl-tRNA synthetases. Mol. Cell 12, 287-294. https://doi.org/10.1016/S1097-2765(03)00280-6

Copyright

Cell Press/Elsevier