title: An archaeal tRNA-synthetase complex that enhances aminoacylation under extreme conditions.
abstract: Aminoacyl-tRNA synthetases (aaRSs) play an integral role in protein synthesis, functioning to attach the correct amino acid with its cognate tRNA molecule. AaRSs are known to associate into higher-order multi-aminoacyl-tRNA synthetase complexes (MSC) involved in archaeal and eukaryotic translation, although the precise biological role remains largely unknown. To gain further insights into archaeal MSCs, possible protein-protein interactions with the atypical Methanothermobacter thermautotrophicus seryl-tRNA synthetase (MtSerRS) were investigated. Yeast two-hybrid analysis revealed arginyl-tRNA synthetase (MtArgRS) as an interacting partner of MtSerRS. Surface plasmon resonance confirmed stable complex formation, with a dissociation constant (K(D)) of 250 nM. Formation of the MtSerRS·MtArgRS complex was further supported by the ability of GST-MtArgRS to co-purify MtSerRS and by coelution of the two enzymes during gel filtration chromatography. The MtSerRS·MtArgRS complex also contained tRNA(Arg), consistent with the existence of a stable ribonucleoprotein complex active in aminoacylation. Steady-state kinetic analyses revealed that addition of MtArgRS to MtSerRS led to an almost 4-fold increase in the catalytic efficiency of serine attachment to tRNA, but had no effect on the activity of MtArgRS. Further, the most pronounced improvements in the aminoacylation activity of MtSerRS induced by MtArgRS were observed under conditions of elevated temperature and osmolarity. These data indicate that formation of a complex between MtSerRS and MtArgRS provides a means by which methanogenic archaea can optimize an early step in translation under a wide range of extreme environmental conditions.
authors.affiliation: Department of Chemistry, Faculty of Science, University of Zagreb, Horvatovac 102a, HR-10000 Zagreb, Croatia.
authors.forename: Vlatka
authors.initials: V
authors.lastname: Godinic-Mikulcic
authors.forename: Jelena
authors.initials: J
authors.lastname: Jaric
authors.forename: Corinne D
authors.initials: CD
authors.lastname: Hausmann
authors.forename: Michael
authors.initials: M
authors.lastname: Ibba
authors.forename: Ivana
authors.initials: I
authors.lastname: Weygand-Durasevic
mesh_terms: D049148:Aminoacylation; D001105:Archaea; D001126:Arginine-tRNA Ligase / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D017013:Methanobacteriaceae / Q000201:enzymology*; D046912:Multiprotein Complexes; D009994:Osmolar Concentration; D014176:Protein Biosynthesis; D025941:Protein Interaction Mapping / Q000379:methods; D012714:Serine-tRNA Ligase / Q000378:metabolism*; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D046912:Multiprotein Complexes; D012714:Serine-tRNA Ligase; D001126:Arginine-tRNA Ligase