In vivo formation of glutamyl-tRNA(Gln) in Escherichia coli by heterologous glutamyl-tRNA synthetases.
Two types of glutamyl-tRNA synthetase exist: the discriminating enzyme (D-GluRS) forms only Glu-tRNA(Glu), while the non-discriminating one (ND-GluRS) also synthesizes Glu-tRNA(Gln), a required intermediate in protein synthesis in many organisms (but not in Escherichia coli). Testing the capacity to complement a thermosensitive E. coli gltX mutant and to suppress an E. coli trpA49 missense mutant we examined the properties of heterologous gltX genes. We demonstrate that while Acidithiobacillus ferrooxidans GluRS1 and Bacillus subtilis Q373R GluRS form Glu-tRNA(Glu), A. ferrooxidans and Helicobacter pylori GluRS2 form Glu-tRNA(Gln) in E. coli in vivo.
Núñez
Harold
H
Programa de Biologi;a Celular y Molecular, ICBM, Facultad de Medicina, Universidad de Chile, Casilla 70086, Santiago 838-0453, Chile.
Lefimil
Claudia
C
Min
Bokkee
B
Söll
Dieter
D
Orellana
Omar
O
D001483:Base Sequence; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005975:Glutamate-tRNA Ligase / Q000235:genetics / Q000378:metabolism*; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D013379:Substrate Specificity
D017931:DNA Primers; D029968:Escherichia coli Proteins; D012346:RNA, Transfer, Amino Acyl; C483187:glutamyl-tRNA(Gln); D005975:Glutamate-tRNA Ligase