The Escherichia coli trmE (mnmE) gene, involved in tRNA modification, codes for an evolutionarily conserved GTPase with unusual biochemical properties

Hugo Cabedo, Fernando Macian-Juan, Magda Villarroya, Juan C. Escudero, Marta Martínez-Vicente, Erwin Knecht, M. Eugenia Armengod

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

The evolutionarily conserved 50K protein of Escherichia coli, encoded by o454, contains a consensus GTP-binding motif. Here we show that 50K is a GTPase that differs extensively from regulatory GTPases such as p21. Thus, 50K exhibits a very high intrinsic GTPase hydrolysis rate, rather low affinity for GTP, and extremely low affinity for GDP. Moreover, it can form self-assemblies. Strikingly, the 17 kDa GTPase domain of 50K conserves the guanine nucleotide-binding and GTPase activities of the intact 50K molecule. Therefore, the structural requirements for GTP binding and GTP hydrolysis by 50K are without precedent and justify a separate classification in the GTPase superfamily. Immunoelectron microscopy reveals that 50K is a cytoplasmic protein partially associated with the inner membrane. We prove that o454 is allelic with trmE, a gene involved in the biosynthesis of the hypermodified nucleoside 5-methylaminomethyl-2-thiouridine, which is found in the wobble position of some tRNAs. Our results demonstrate that 50K is essential for viability depending on the genetic background. We propose that combination of mutations affecting the decoding process, which separately do not reveal an obvious defect in growth, can give rise to lethal phenotypes, most likely due to synergism.

Original languageEnglish (US)
Pages (from-to)7063-7076
Number of pages14
JournalEMBO Journal
Volume18
Issue number24
StatePublished - Dec 15 1999
Externally publishedYes

Fingerprint

GTP Phosphohydrolases
Transfer RNA
Escherichia coli
Genes
Guanosine Triphosphate
Hydrolysis
Guanine Nucleotides
Immunoelectron Microscopy
Escherichia coli Proteins
Biosynthesis
Nucleosides
Self assembly
Decoding
Microscopic examination
Membranes
Phenotype
Defects
Mutation
Molecules
Growth

Keywords

  • GTPase
  • Synthetic lethality
  • Translation
  • tRNA modification

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Cabedo, H., Macian-Juan, F., Villarroya, M., Escudero, J. C., Martínez-Vicente, M., Knecht, E., & Armengod, M. E. (1999). The Escherichia coli trmE (mnmE) gene, involved in tRNA modification, codes for an evolutionarily conserved GTPase with unusual biochemical properties. EMBO Journal, 18(24), 7063-7076.

The Escherichia coli trmE (mnmE) gene, involved in tRNA modification, codes for an evolutionarily conserved GTPase with unusual biochemical properties. / Cabedo, Hugo; Macian-Juan, Fernando; Villarroya, Magda; Escudero, Juan C.; Martínez-Vicente, Marta; Knecht, Erwin; Armengod, M. Eugenia.

In: EMBO Journal, Vol. 18, No. 24, 15.12.1999, p. 7063-7076.

Research output: Contribution to journalArticle

Cabedo, H, Macian-Juan, F, Villarroya, M, Escudero, JC, Martínez-Vicente, M, Knecht, E & Armengod, ME 1999, 'The Escherichia coli trmE (mnmE) gene, involved in tRNA modification, codes for an evolutionarily conserved GTPase with unusual biochemical properties', EMBO Journal, vol. 18, no. 24, pp. 7063-7076.
Cabedo, Hugo ; Macian-Juan, Fernando ; Villarroya, Magda ; Escudero, Juan C. ; Martínez-Vicente, Marta ; Knecht, Erwin ; Armengod, M. Eugenia. / The Escherichia coli trmE (mnmE) gene, involved in tRNA modification, codes for an evolutionarily conserved GTPase with unusual biochemical properties. In: EMBO Journal. 1999 ; Vol. 18, No. 24. pp. 7063-7076.
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