A Freestanding Proofreading Domain is Required for Protein Synthesis Quality Control in Archaea

Authors

Dragana Korencic, Yale University
Ivan Ahel, Yale University
James Schelert, University of Nebraska - Lincoln
Mike Sacher, Yale University
Benfang Ruan, Yale University
Constantinos Stathopoulos, Yale University
Paul Blum, University of Nebraska-Lincoln
Michael Ibba, Chapman UniversityFollow
Dieter Söll, Yale University

Document Type

Article

Publication Date

7-13-2004

Abstract

Threonyl-tRNA synthetase (ThrRS) participates in protein synthesis quality control by selectively editing the misacylated species Ser-tRNA^Thr. In bacteria and eukaryotes the editing function of ThrRS resides in a highly conserved N-terminal domain distant from the active site. Most archaeal ThrRS proteins are devoid of this editing domain, suggesting evolutionary divergence of quality-control mechanisms. Here we show that archaeal editing of Ser-tRNA^Thr is catalyzed by a domain unrelated to, and absent from, bacterial and eukaryotic ThrRSs. Despite the lack of sequence homology, the archaeal and bacterial editing domains are both reliant on a pair of essential histidine residues suggestive of a common catalytic mechanism. Whereas the archaeal editing module is most commonly part of full-length ThrRS, several crenarchaeal species contain individual genes encoding the catalytic (ThrRS-cat) and editing domains (ThrRS-ed). Sulfolobus solfataricus ThrRS-cat was shown to synthesize both Thr-tRNA^Thr and Ser-tRNA^Thr and to lack editing activity against Ser-tRNA^Thr. In contrast, ThrRS-ed lacks aminoacylation activity but can act as an autonomous protein in trans to hydrolyze specifically Ser-tRNA^Thr, or it can be fused to ThrRS-cat to provide the same function in cis. Deletion analyses indicate that ThrRS-ed is dispensable for growth of S. solfataricus under standard conditions but is required for normal growth in media with elevated serine levels. The growth phenotype of the ThrRS-ed deletion strain suggests that retention of the discontinuous ThrRS quaternary structure relates to specific physiological requirements still evident in certain Archaea.

Comments

This article was originally published in Proceedings of the National Academy of Sciences of the United States of America, volume 101, in 2004. https://doi.org/10.1073/pnas.0403926101

Recommended Citation

Korencic, D., Ahel, I., Schelert, J., Sacher, M., Ruan, B., Stathopoulos, C., Blum, P., Ibba, M. and Söll, D. (2004) A freestanding proofreading domain is required for protein synthesis quality control in Archaea. Proc. Natl. Acad. Sci. USA 101, 10260-10265. https://doi.org/10.1073/pnas.0403926101

Copyright

The National Academy of Sciences