title: Identification of antituberculosis agents that target ribosomal protein interactions using a yeast two-hybrid system.
abstract: Mycobacterium tuberculosis kills about 2 million people annually and antibiotic resistance is a cause of increased mortality. Therefore, development of new antituberculosis drugs is urgent for the control of widespread tuberculosis infections. For this purpose, we performed an innovative screen to identify new agents that disrupt the function of ribosomes in M. tuberculosis. Two bacterial ribosomal proteins L12 and L10 interact with each other and constitute the stalk of the 50S ribosomal subunit, which recruits initiation and elongation factors (EFs) during translation. Therefore, the L12-L10 interaction should be essential for ribosomal function and protein synthesis. We established a yeast two-hybrid system to identify small molecules that block the interaction between L12 and L10 proteins from M. tuberculosis. Using this system, we identified two compounds T766 and T054 that show strong bactericidal activity against tuberculosis but with low toxicity to mice and other bacterial strains. Moreover, using surface plasmon resonance (SPR) assay, we have demonstrated that these compounds bind specifically to L12 to disrupt L12-L10 interaction. Overproduction of L12 protein, but not L10, lowers the antibacterial activity of T766 and T054, indicating that the ribosome is likely the cellular target. Therefore, our data demonstrate that this yeast two-hybrid system is a useful tool to identify unique antituberculosis agents targeting the ribosomal protein L12-L10 interaction.
authors.affiliation: Key Laboratory of Biotechnology of Antibiotics of Ministry of Health, Institute of Medicinal Biotechnology,  Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050,  China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lin
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Yuanjun
authors.initials: Y
authors.lastname: Zhu
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Yongzhen
authors.initials: Y
authors.lastname: Li
authors.forename: Xiao
authors.initials: X
authors.lastname: Liu
authors.forename: Wei
authors.initials: W
authors.lastname: Jiang
authors.forename: Shishan
authors.initials: S
authors.lastname: Yu
authors.forename: Xue-Fu
authors.initials: XF
authors.lastname: You
authors.forename: Chunling
authors.initials: C
authors.lastname: Xiao
authors.forename: Bin
authors.initials: B
authors.lastname: Hong
authors.forename: Yanchang
authors.initials: Y
authors.lastname: Wang
authors.forename: Jian-Dong
authors.initials: JD
authors.lastname: Jiang
authors.forename: Shuyi
authors.initials: S
authors.lastname: Si
mesh_terms: D000995:Antitubercular Agents / Q000378:metabolism / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000254:growth & development; D012269:Ribosomal Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D020349:Surface Plasmon Resonance; D020798:Two-Hybrid System Techniques
chemical_list: D000995:Antitubercular Agents; D012269:Ribosomal Proteins