A Conserved Lysine in an Ion-Pair with a Catalytic Glutamate Is Critical for U-to-C RNA Editing but Restricts C-to-U RNA Editing DOI

Skellie O. Chun,

Elvin T. Garcia,

Marcela Rejas

et al.

Biochemistry, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 9, 2024

Plants make pyrimidine base substitutions in organellar mRNAs through the action of sequence-specific nuclear-encoded enzymes. Pentatricopeptide repeat (PPR) proteins are essential for ensuring specificity, while enzymatic DYW domain is often present at C-terminus a PPR protein and dependent on variant possessing C-to-U and/or U-to-C RNA editing activities. Expression exogenous DYW-KP enzymes bacteria leads to modification RNAs suggestive changes. The modified could only be purified from interphase an acidic guanidinium thiocyanate-phenol-chloroform experiment. It was projected that stable RNA-enzyme cross-links form lysyl attack. In this study, examined dual U-to-C/C-to-U enzyme KP6 with conserved lysine residues substituted by alanine. A single found and, based crystal structures domains, would likely active site. Crystal also suggest can potentially ion pair catalytic glutamate critical editing. Mutation alanine greatly stimulated KP6. ∼319 Da adduct observed not detected U-to-C-deficient point mutant This work establishes role specifically activity but highlights secondary modulating formation inhibitory glutamate.

Language: Английский

Phylogenetic and Structural Analysis of Hydra ADAR DOI Creative Commons
Xander E. Wilcox, Heng Zhang, Jasmine L. Mah

et al.

Archives of Biochemistry and Biophysics, Journal Year: 2025, Volume and Issue: unknown, P. 110353 - 110353

Published: Feb. 1, 2025

Language: Английский

Citations

0
A Conserved Lysine in an Ion-Pair with a Catalytic Glutamate Is Critical for U-to-C RNA Editing but Restricts C-to-U RNA Editing DOI

Skellie O. Chun,

Elvin T. Garcia,

Marcela Rejas

et al.

Biochemistry, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 9, 2024

Plants make pyrimidine base substitutions in organellar mRNAs through the action of sequence-specific nuclear-encoded enzymes. Pentatricopeptide repeat (PPR) proteins are essential for ensuring specificity, while enzymatic DYW domain is often present at C-terminus a PPR protein and dependent on variant possessing C-to-U and/or U-to-C RNA editing activities. Expression exogenous DYW-KP enzymes bacteria leads to modification RNAs suggestive changes. The modified could only be purified from interphase an acidic guanidinium thiocyanate-phenol-chloroform experiment. It was projected that stable RNA-enzyme cross-links form lysyl attack. In this study, examined dual U-to-C/C-to-U enzyme KP6 with conserved lysine residues substituted by alanine. A single found and, based crystal structures domains, would likely active site. Crystal also suggest can potentially ion pair catalytic glutamate critical editing. Mutation alanine greatly stimulated KP6. ∼319 Da adduct observed not detected U-to-C-deficient point mutant This work establishes role specifically activity but highlights secondary modulating formation inhibitory glutamate.

Language: Английский

Citations

1