Antimicrobial resistance is a major health threat as it limits treatment options for infection. At the forefront of this serious issue is , a Gram-negative opportunistic pathogen that exhibits the remarkable ability to resist antibiotics through multiple mechanisms. As bacterial ribosomes represent a target for multiple distinct classes of existing antimicrobial agents, we here use single-particle cryo-electron microscopy (cryo-EM) to elucidate five different structural states of the ribosome, including the 70S, 50S, and 30S forms. We also determined interparticle motions of the 70S ribosome in different tRNA bound states using three-dimensional (3D) variability analysis. Together, our structural data further our understanding of the ribosome from and other Gram-negative pathogens and will enable structure-based drug discovery to combat antibiotic-resistant bacterial infections. is a severe nosocomial threat largely due to its intrinsic antibiotic resistance and remarkable ability to acquire new resistance determinants. The bacterial ribosome serves as a major target for modern antibiotics and the design of new therapeutics. Here, we present cryo-EM structures of the 70S ribosome, revealing several unique species-specific structural features that may facilitate future drug development to combat this recalcitrant bacterial pathogen.
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http://dx.doi.org/10.1128/mBio.03117-19 | DOI Listing |
Biosci Biotechnol Biochem
December 2024
Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu-shi, Toyama, Japan.
ABCF proteins (ABCFs) are key components of prokaryotic translation systems, resolving ribosomal stalling. These ATPases contain two ATPase domains and interdomain linker, the length and composition of which are key determinants of their function. Antibiotic resistance ABCF (ARE-ABCFs) proteins, counteract ribosome-targeting antibiotics by binding to the E site of the 70S ribosome, promoting drug dissociation.
View Article and Find Full Text PDFJ Biol Chem
December 2024
Institut für Zytobiologie im Zentrum für Synthetische Mikrobiologie SynMikro, Philipps-Universität Marburg, Karl-von-Frisch-Str. 14, 35032 Marburg, Germany. Electronic address:
Mitochondria synthesize only a small set of their proteins on endogenous mitoribosomes. These particles differ in structure and composition from both their bacterial 70S ancestors and cytosolic 80S ribosomes. Recently published high resolution structures of the human mitoribosome revealed the presence of three [2Fe-2S] clusters in the small and large subunits.
View Article and Find Full Text PDFBiochim Biophys Acta Gen Subj
January 2025
Department of Biochemistry and Biophysics, University of Kalyani, Kalyani 741 235, West Bengal, India. Electronic address:
Background: In earlier communications we reported about nanonization of the antibiotic tetracycline (Tet) by entrapping it within the biocompatible and highly membrane penetrating nano-carrier molecule - calcium phosphate nanoparticle (CPNP). The synthesized Tet-CPNP killed different Tet-resistant bacteria in vitro as well as in vivo (in mice). Moreover, such nanonized tetracycline had bactericidal mode of action, in contrast to bacteriostatic mode of action of bulk tetracycline.
View Article and Find Full Text PDFbioRxiv
November 2024
Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, 30322, USA.
Loss of ribosomal RNA (rRNA) modifications incorporated by the intrinsic methyltransferase TlyA results in reduced sensitivity to tuberactinomycin antibiotics such as capreomycin. However, the mechanism by which rRNA methylation alters drug binding, particularly at the distant but functionally more important site in 23S rRNA Helix 69 (H69), is currently unknown. We determined high-resolution cryo-electron microscopy structures of the 70S ribosome with or without the two ribose 2'-O-methyl modifications incorporated by TlyA.
View Article and Find Full Text PDFbioRxiv
October 2024
Department of Microbiology, Cornell University, Ithaca, NY 14853.
Polyproline motifs are essential structural features of many proteins, and recent evidence suggests that EF-P is one of several factors that facilitate their translation. For example, YfmR was recently identified as a protein that prevents ribosome stalling at proline-containing sequences in the absence of EF-P. Here, we show that the YebC-family protein YebC2 (formerly YeeI) functions as a translation factor in that resolves ribosome stalling at polyprolines.
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