Structural and functional characterization of the bacterial translocation inhibitor GE82832

Brandi, Letizia; Maffioli, Sonia; Donadio, Stefano; Quaglia, Fabio; Sette, Marco; Milón, Pohl; Gualerzi, Claudio O.; Fabbretti, Attilio

doi:10.1016/j.febslet.2012.07.040

Cited by 23 publications

(32 citation statements)

References 38 publications

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“…Indeed, screening systems to identify compounds that preferentially inhibit the initiation phase have proved successful [51,52,53]. In addition, our IF3 DL -30S reporter assay can provide novel aspects of the inhibiting mechanism of known 30S-binding drugs.…”

Section: Discussionmentioning

confidence: 99%

Conformational Response of 30S-bound IF3 to A-Site Binders Streptomycin and Kanamycin

et al. 2016

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Aminoglycoside antibiotics are widely used to treat infectious diseases. Among them, streptomycin and kanamycin (and derivatives) are of importance to battle multidrug-resistant (MDR) Mycobacterium tuberculosis. Both drugs bind the small ribosomal subunit (30S) and inhibit protein synthesis. Genetic, structural, and biochemical studies indicate that local and long-range conformational rearrangements of the 30S subunit account for this inhibition. Here, we use intramolecular FRET between the C- and N-terminus domains of the flexible IF3 to monitor real-time perturbations of their binding sites on the 30S platform. Steady and pre-steady state binding experiments show that both aminoglycosides bring IF3 domains apart, promoting an elongated state of the factor. Binding of Initiation Factor IF1 triggers closure of IF3 bound to the 30S complex, while both aminoglycosides revert the IF1-dependent conformation. Our results uncover dynamic perturbations across the 30S subunit, from the A-site to the platform, and suggest that both aminoglycosides could interfere with prokaryotic translation initiation by modulating the interaction between IF3 domains with the 30S platform.

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Section: Discussionmentioning

confidence: 99%

Conformational Response of 30S-bound IF3 to A-Site Binders Streptomycin and Kanamycin

et al. 2016

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“…Meanwhile, several lines of evidence suggest that a large scale conformational change of EF-G occurs during translocation (Agrawal et al, 1999; Bulkley et al, 2014; Munro et al, 2010; Salsi et al, 2014; Stark et al, 2000; Wang et al, 2007), including a recent study of the antibiotic dityromycin (Bulkley et al, 2014). Dityromycin was shown to block EF-G-mediated tRNA translocation without affecting the binding of EF-G to the ribosome (Brandi et al, 2006; Brandi et al, 2012). The crystal structure of dityromycin in complex with the ribosome shows that dityromycin binds to ribosomal protein S12 (Bulkley et al, 2014), a position that would severely overlap with domain III of EF-G in the elongated form, thereby indicating the necessity for substantial domain rearrangements in EF-G on the PRE ribosome.…”

Section: Introductionmentioning

confidence: 99%

Conformational Changes of Elongation Factor G on the Ribosome during tRNA Translocation

Lin

Gagnon

Bulkley

et al. 2015

Cell

123

153

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Summary The universally conserved GTPase elongation factor G (EF-G) catalyzes the translocation of transfer RNA (tRNA) and messenger RNA (mRNA) on the ribosome after peptide bond formation. Despite numerous studies suggesting that EF-G undergoes extensive conformational rearrangements during translocation, high resolution structures exist for essentially only one conformation of EF-G in complex with the ribosome. Here, we report four atomic resolution crystal structures of EF-G bound to the ribosome programmed in the pre- and post-translocational states and to the ribosome trapped by the antibiotic dityromycin. We observe a previously unseen conformation of EF-G in the pretranslocation complex, which is independently captured by dityromycin on the ribosome. Our structures provide insights into the conformational space that EF-G samples on the ribosome and reveal that tRNA translocation on the ribosome is facilitated by a structural transition of EF-G from a compact to an elongated conformation, which can be prevented by the antibiotic dityromycin.

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“…The antibiotic dityromycin has also been shown to block EF-G associated translocation 107 . The crystal structure of dityromycin in complex with the ribosome reveals that it binds to ribosomal protein S12 and would clash with domain III of EF-G in its elongated form 108 (Fig.…”

Section: Structural Comparison Of Isolated Ef-g Ef4 and Bipamentioning

confidence: 99%

Similarity and diversity of translational GTPase factors EF-G, EF4, and BipA: From structure to function

et al. 2016

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EF-G, EF4, and BipA are members of the translation factor family of GTPases with a common ribosome binding mode and GTPase activation mechanism. However, topological variations of shared as well as unique domains ensure different roles played by these proteins during translation. Recent X-ray crystallography and cryo-electron microscopy studies have revealed the structural basis for the involvement of EF-G domain IV in securing the movement of tRNAs and mRNA during translocation as well as revealing how the unique C-terminal domains of EF4 and BipA interact with the ribosome and tRNAs contributing to the regulation of translation under certain conditions. EF-G, EF-4, and BipA are intriguing examples of structural variations on a common theme that results in diverse behavior and function. Structural studies of translational GTPase factors have been greatly facilitated by the use of antibiotics, which have revealed their mechanism of action.

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Structural and functional characterization of the bacterial translocation inhibitor GE82832

Cited by 23 publications

References 38 publications

Conformational Response of 30S-bound IF3 to A-Site Binders Streptomycin and Kanamycin

Conformational Response of 30S-bound IF3 to A-Site Binders Streptomycin and Kanamycin

Conformational Changes of Elongation Factor G on the Ribosome during tRNA Translocation

Similarity and diversity of translational GTPase factors EF-G, EF4, and BipA: From structure to function

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