Document Type
Article
Publication Date
3-19-2021
Publication Title
iScience
Abstract
Aminoacyl-tRNA synthetases (AARS) participate in decoding the genome by catalyzing conjugation of amino acids to their cognate tRNAs. During evolution, biochemical and environmental conditions markedly influenced the sequence and structure of the 20 AARSs, revealing adaptations dictating canonical and orthogonal activities. Here, we investigate the function of the appended Zn2+-binding domain (ZBD) in the bifunctional AARS, glutamyl-prolyl-tRNA synthetase (GluProRS). We developed GluProRS mutant mice by CRISPR-Cas9 with a deletion of 29 C-terminal amino acids, including two of four Zn2+-coordinating cysteines. Homozygous ZBD mutant mice die before embryonic day 12.5, but heterozygous mice are healthy. ZBD disruption profoundly reduces GluProRS canonical function by dual mechanisms: it induces rapid proteasomal degradation of the protein and inhibits ProRS aminoacylation activity, likely by suboptimal positioning of ATP in the spatially adjacent catalytic domain. Collectively, our studies reveal the ZBD as a critical determinant of ProRS activity and GluProRS stability in vitro and in vivo.
Recommended Citation
Vasu, Kommireddy; Ramachandiran, Iyappan; Terenzi, Fulvia; Khan, Debjit; China, Arnab; Khan, Krishnendu; Chechi, Aayushi; Baleanu-Gogonea, Camelia; Gogonea, Valentin; and Fox, Paul L., "The Zinc-binding Domain of Mammalian prolyl-tRNA synthetase is Indispensable for Catalytic Activity and Organism Viability" (2021). Chemistry Faculty Publications. 557.
https://engagedscholarship.csuohio.edu/scichem_facpub/557
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
DOI
10.1016/j.isci.2021.102215
Version
Publisher's PDF
Volume
24
Issue
3
Comments
This project was supported by NIH grants P01 HL076491, R01 DK123236, and R01 AG067146 (to P.L.F.), by aClinical and Translational Science Collaborative (CTSC) Core Utilization Pilot Grant Award from Case West-ern Reserve University School of Medicine (to P.L.F.), and by American Heart Association (AHA) Postdoc-toral Fellowships, Nationwide (19POST34380725 to I.R. and 19POST34380687 to D.K.)