Publications

• Role of D-aminoacyl-tRNA deacylase beyond chiral proofreading as a cellular defense against glycine mischarging by AlaRS.

  Pawar K.I., Suma K., Seenivasan A., Kuncha S.K., Routh S.B., Kruparani S.P., Sankaranarayanan R.

  Strict L-chiral rejection through Gly-<i>cis</i>Pro motif during chiral proofreading underlies the inability of D-aminoacyl-tRNA deacylase (DTD) to discriminate between D-amino acids and achiral glycine. The consequent Gly-tRNA^Gly 'misediting paradox' is resolved by EF-Tu in the cell. H ... >> More

  Strict L-chiral rejection through Gly-<i>cis</i>Pro motif during chiral proofreading underlies the inability of D-aminoacyl-tRNA deacylase (DTD) to discriminate between D-amino acids and achiral glycine. The consequent Gly-tRNA^Gly 'misediting paradox' is resolved by EF-Tu in the cell. Here, we show that DTD's active site architecture can efficiently edit mischarged Gly-tRNA^Ala species four orders of magnitude more efficiently than even AlaRS, the only ubiquitous cellular checkpoint known for clearing the error. Also, DTD knockout in AlaRS editing-defective background causes pronounced toxicity in <i>Escherichia coli</i> even at low-glycine levels which is alleviated by alanine supplementation. We further demonstrate that DTD positively selects the universally invariant tRNA^Ala-specific G3•U70. Moreover, DTD's activity on non-cognate Gly-tRNA^Ala is conserved across all bacteria and eukaryotes, suggesting DTD's key cellular role as a glycine deacylator. Our study thus reveals a hitherto unknown function of DTD in cracking the universal mechanistic dilemma encountered by AlaRS, and its physiological importance. << Less

  Elife 6:0-0(2017) [PubMed] [EuropePMC]

• Unexpected enzymatic function of an ancient nucleic acid-binding fold.

  Watkins R.R., Bockelman S., Vradi A., Grabarkewitz K., Pyun A., Stark J., Wysocki V.H., Alfonzo J.D., Musier-Forsyth K.

  Aminoacyl-tRNA synthetases (ARSs) are indispensable for all living organisms and their associated aminoacyl-tRNA editing domains ensure the fidelity of translation. In eukaryotes, ARSs form a multi-aminoacyl-tRNA synthetase complex (MSC), which is assembled together with several nonsynthetase scaf ... >> More

  Aminoacyl-tRNA synthetases (ARSs) are indispensable for all living organisms and their associated aminoacyl-tRNA editing domains ensure the fidelity of translation. In eukaryotes, ARSs form a multi-aminoacyl-tRNA synthetase complex (MSC), which is assembled together with several nonsynthetase scaffolding proteins. The MSC found in Trypanosoma brucei (Tb) includes two proteins with oligosaccharide/oligonucleotide-binding (OB) folds-MSC-associated protein 1 (MCP1) and MCP2-and one known trans-editing factor, MCP3, an Ala-tRNA deacylase. The activity of MCP1 was unexplored until now. Our study shows that recombinantly-expressed and purified MCP1 also deacylates Ala-tRNAs despite lacking known tRNA-editing domain homology. Domain deletion studies reveal that the OB-fold houses the catalytic pocket and mutation of any one of three conserved OB-fold residues (K326, R331, S335) abolishes activity. Assays with Saccharomyces cerevisiae Arc1p reveal that MCP1's deacylation activity is conserved across organisms. This discovery explains the 3' CCA-end binding activity of this protein family and uncovers an ancient nucleic acid binding domain's unexpected enzymatic function. << Less

  Nucleic Acids Res. 53:0-0(2025) [PubMed] [EuropePMC]

  This publication is cited by 3 other entries.