Identification of Anti-Gram-negative Bacteria Agents Targeting the Interaction Between Ribosomal Proteins L12 and L10

Overview

Journal Acta Pharm Sin B

Publisher Elsevier

Specialty Pharmacology

Date 2018 Sep 25

PMID 30245964

Citations 4

Authors

Weiwei Wang

Chao Liu

Ningyu Zhu

Yuan Lin

Jiandong Jiang

Yanchang Wang

Yan Li

Shuyi Si

Affiliations

Soon will be listed here.

Abstract

Gram-negative bacteria have become the main pathogens and cause serious clinical problems with increased morbidity and mortality. However, the slow discovery of new antimicrobial agents is unable to meet the need for the treatment of bacterial infections caused by drug-resistant strains. The interaction of L12 and L10 is essential for ribosomal function and protein synthesis. In this study, a yeast two-hybrid system was established to successfully detect the interaction between L12 and L10 proteins from gram-negative bacteria , which allows us to screen compounds that specifically disrupt this interaction. With this system, we identified two compounds IMB-84 and IMB-87 that block L12-L10 interaction and show bactericidal activity against . We used glutathione--transferase (GST) pull-down and surface plasmon resonance (SPR) assays to demonstrate that these compounds disrupt L12-L10 interaction and the target of compounds was further confirmed by the overexpression of target proteins. Moreover, protein synthesis and elongation factor G-dependent GTPase activities are inhibited by two compounds. Therefore, we have identified two antibacterial agents that disrupt L12-L10 interaction by using yeast two-hybrid system.

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References

Savelsbergh A, Mohr D, Wilden B, Wintermeyer W, Rodnina M . Stimulation of the GTPase activity of translation elongation factor G by ribosomal protein L7/12. J Biol Chem. 2000; 275(2):890-4. DOI: 10.1074/jbc.275.2.890. View

Zengel J, Archer R, Lindahl L . The nucleotide sequence of the Escherichia coli fus gene, coding for elongation factor G. Nucleic Acids Res. 1984; 12(4):2181-92. PMC: 318650. DOI: 10.1093/nar/12.4.2181. View

Li B, Fields S . Identification of mutations in p53 that affect its binding to SV40 large T antigen by using the yeast two-hybrid system. FASEB J. 1993; 7(10):957-63. DOI: 10.1096/fasebj.7.10.8344494. View

Wilson D, Nierhaus K . Ribosomal proteins in the spotlight. Crit Rev Biochem Mol Biol. 2005; 40(5):243-67. DOI: 10.1080/10409230500256523. View

Hanes J, Freudenstein J, Rapp G, Scheit K . Construction of a plasmid containing the complete coding region of human elongation factor 2. Biol Chem Hoppe Seyler. 1992; 373(4):201-4. DOI: 10.1515/bchm3.1992.373.1.201. View

Gietz R, Schiestl R, Willems A, Woods R . Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast. 1995; 11(4):355-60. DOI: 10.1002/yea.320110408. View

Baum E, Crespo-Carbone S, Foleno B, Simon L, Guillemont J, Macielag M . MurF inhibitors with antibacterial activity: effect on muropeptide levels. Antimicrob Agents Chemother. 2009; 53(8):3240-7. PMC: 2715636. DOI: 10.1128/AAC.00166-09. View

Lin Y, Li Y, Zhu Y, Zhang J, Li Y, Liu X . Identification of antituberculosis agents that target ribosomal protein interactions using a yeast two-hybrid system. Proc Natl Acad Sci U S A. 2012; 109(43):17412-7. PMC: 3491523. DOI: 10.1073/pnas.1110271109. View

Bischoff L, Berninghausen O, Beckmann R . Molecular basis for the ribosome functioning as an L-tryptophan sensor. Cell Rep. 2014; 9(2):469-75. DOI: 10.1016/j.celrep.2014.09.011. View

10.

Studier F . Protein production by auto-induction in high density shaking cultures. Protein Expr Purif. 2005; 41(1):207-34. DOI: 10.1016/j.pep.2005.01.016. View

11.

Vincent J, Rello J, Marshall J, Silva E, Anzueto A, Martin C . International study of the prevalence and outcomes of infection in intensive care units. JAMA. 2009; 302(21):2323-9. DOI: 10.1001/jama.2009.1754. View

12.

Savelsbergh A, Mohr D, Kothe U, Wintermeyer W, Rodnina M . Control of phosphate release from elongation factor G by ribosomal protein L7/12. EMBO J. 2005; 24(24):4316-23. PMC: 1356325. DOI: 10.1038/sj.emboj.7600884. View

13.

Yonath A . Antibiotics targeting ribosomes: resistance, selectivity, synergism and cellular regulation. Annu Rev Biochem. 2005; 74:649-79. DOI: 10.1146/annurev.biochem.74.082803.133130. View

14.

Wellington E, Boxall A, Cross P, Feil E, Gaze W, Hawkey P . The role of the natural environment in the emergence of antibiotic resistance in gram-negative bacteria. Lancet Infect Dis. 2013; 13(2):155-65. DOI: 10.1016/S1473-3099(12)70317-1. View

15.

Gonzalo P, Reboud J . The puzzling lateral flexible stalk of the ribosome. Biol Cell. 2003; 95(3-4):179-93. DOI: 10.1016/s0248-4900(03)00034-0. View

16.

Lin Y, Li Y, Zhu N, Han Y, Jiang W, Wang Y . The antituberculosis antibiotic capreomycin inhibits protein synthesis by disrupting interaction between ribosomal proteins L12 and L10. Antimicrob Agents Chemother. 2014; 58(4):2038-44. PMC: 4023752. DOI: 10.1128/AAC.02394-13. View

17.

Nambiar S, Laessig K, Toerner J, Farley J, Cox E . Antibacterial drug development: challenges, recent developments, and future considerations. Clin Pharmacol Ther. 2014; 96(2):147-9. DOI: 10.1038/clpt.2014.116. View

18.

Pettersson I . Studies on the RNA and protein binding sites of the E. coli ribosomal protein L10. Nucleic Acids Res. 1979; 6(7):2637-46. PMC: 327877. DOI: 10.1093/nar/6.7.2637. View

19.

Chaudhary A . A review of global initiatives to fight antibiotic resistance and recent antibiotics׳ discovery. Acta Pharm Sin B. 2016; 6(6):552-556. PMC: 5071619. DOI: 10.1016/j.apsb.2016.06.004. View

20.

Brown E, Wright G . Antibacterial drug discovery in the resistance era. Nature. 2016; 529(7586):336-43. DOI: 10.1038/nature17042. View