Specific Recognition of a Minimal Model of Aminoacylated tRNA by the Elongation Factor Tu of Bacterial Protein Biosynthesis

Hsp33, a prokaryotic redox-regulated holding chaperone, has been recently identified to be able to exhibit an unfoldase and aggregase activity against elongation factor Tu (EF-Tu) in its reduced state. In this study, we investigated the effect of elongation factor Ts (EF-Ts) and trigger factor (TF) on Hsp33-mediated EF-Tu unfolding and aggregation using gel filtration, light scattering, circular dichroism, and isothermal titration calorimetry. We found that EF-Tu unfolding and subsequent aggregation induced by Hsp33 were evident even in its complex state with EF-Ts, which enhanced EF-Tu stability. In addition, although TF alone had no substantial effect on the stability of EF-Tu, it markedly amplified the Hsp33-mediated EF-Tu unfolding and aggregation. Collectively, the present results constitute the first example of synergistic unfoldase/aggregase activity of molecular chaperones and suggest that the stability of EF-Tu is modulated by a sophisticated network of molecular chaperones to regulate protein biosynthesis in cells under stress conditions.

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Kirromycin, an Inhibitor of Protein Biosynthesis that Acts on Elongation Factor Tu

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.71.12.4910 ◽

1974 ◽

Vol 71 (12) ◽

pp. 4910-4914 ◽

Cited By ~ 116

Author(s):

H. Wolf ◽

G. Chinali ◽

A. Parmeggiani

Keyword(s):

Protein Biosynthesis ◽

Elongation Factor ◽

Elongation Factor Tu

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Fluorescence characterization of the interaction of various transfer RNA species with elongation factor Tu.cntdot.GTP: evidence for a new functional role for elongation factor Tu in protein biosynthesis

Biochemistry ◽

10.1021/bi00470a002 ◽

1990 ◽

Vol 29 (18) ◽

pp. 4268-4277 ◽

Cited By ~ 59

Author(s):

Fabiola Janiak ◽

V. Ann Dell ◽

Julie K. Abrahamson ◽

Bonnie S. Watson ◽

David L. Miller ◽

...

Keyword(s):

Functional Role ◽

Protein Biosynthesis ◽

Elongation Factor ◽

Transfer Rna ◽

Elongation Factor Tu

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Elongation Factor Tu Mutants Expand Amino Acid Tolerance of Protein Biosynthesis System

Journal of the American Chemical Society ◽

10.1021/ja075557u ◽

2007 ◽

Vol 129 (46) ◽

pp. 14458-14462 ◽

Cited By ~ 77

Author(s):

Yoshio Doi ◽

Takashi Ohtsuki ◽

Yoshihiro Shimizu ◽

Takuya Ueda ◽

Masahiko Sisido

Keyword(s):

Amino Acid ◽

Protein Biosynthesis ◽

Elongation Factor ◽

Acid Tolerance ◽

Elongation Factor Tu

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In VitroandIn VivoActivities of Novel, Semisynthetic Thiopeptide Inhibitors of Bacterial Elongation Factor Tu

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00582-11 ◽

2011 ◽

Vol 55 (11) ◽

pp. 5277-5283 ◽

Cited By ~ 18

Author(s):

J. A. Leeds ◽

M. J. LaMarche ◽

J. T. Brewer ◽

S. M. Bushell ◽

G. Deng ◽

...

Keyword(s):

Bacterial Infections ◽

Elongation Factor ◽

Infection Model ◽

Elongation Factor Tu ◽

Bacterial Protein ◽

Preclinical Development ◽

Gram Positive ◽

Content Type

ABSTRACTRecently, we identified aminothiazole derivatives of GE2270 A. These novel semisynthetic congeners, like GE2270 A, target the essential bacterial protein elongation factor Tu (EF-Tu). Medicinal chemistry optimization of lead molecules led to the identification of preclinical development candidates 1 and 2. These cycloalklycarboxylic acid derivatives show activity against difficult to treat Gram-positive pathogens and demonstrate increased aqueous solubility compared to GE2270 A. We describe here thein vitroandin vivoactivities of compounds 1 and 2 compared to marketed antibiotics. Compounds 1 and 2 were potent against clinical isolates of methicillin-resistantStaphylococcus aureusand vancomycin-resistant enterococci (MIC90≤ 0.25 μg/ml) but weaker against the streptococci (MIC90≥ 4 μg/ml). Like GE2270 A, the derivatives inhibited bacterial protein synthesis and selected for spontaneous loss of susceptibility via mutations in thetufgene, encoding EF-Tu. The mutants were not cross-resistant to other antibiotic classes. In a mouse systemic infection model, compounds 1 and 2 protected mice from lethalS. aureusinfections with 50% effective doses (ED50) of 5.2 and 4.3 mg/kg, respectively. Similarly, compounds 1 and 2 protected mice from lethal systemicE. faecalisinfections with ED50of 0.56 and 0.23 mg/kg, respectively. In summary, compounds 1 and 2 are activein vitroandin vivoactivity against difficult-to-treat Gram-positive bacterial infections and represent a promising new class of antibacterials for use in human therapy.

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