title: Pyrazinamide inhibits trans-translation in Mycobacterium tuberculosis.
abstract: Pyrazinamide (PZA) is a first-line tuberculosis drug that plays a unique role in shortening the duration of tuberculosis chemotherapy. PZA is hydrolyzed intracellularly to pyrazinoic acid (POA) by pyrazinamidase (PZase, encoded by pncA), an enzyme frequently lost in PZA-resistant strains, but the target of POA in Mycobacterium tuberculosis has remained elusive. Here, we identify a previously unknown target of POA as the ribosomal protein S1 (RpsA), a vital protein involved in protein translation and the ribosome-sparing process of trans-translation. Three PZA-resistant clinical isolates without pncA mutation harbored RpsA mutations. RpsA overexpression conferred increased PZA resistance, and we confirmed that POA bound to RpsA (but not a clinically identified ΔAla mutant) and subsequently inhibited trans-translation rather than canonical translation. Trans-translation is essential for freeing scarce ribosomes in nonreplicating organisms, and its inhibition may explain the ability of PZA to eradicate persisting organisms.
authors.affiliation: W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA.
authors.forename: Wanliang
authors.initials: W
authors.lastname: Shi
authors.forename: Xuelian
authors.initials: X
authors.lastname: Zhang
authors.forename: Xin
authors.initials: X
authors.lastname: Jiang
authors.forename: Haiming
authors.initials: H
authors.lastname: Yuan
authors.forename: Jong Seok
authors.initials: JS
authors.lastname: Lee
authors.forename: Clifton E
authors.initials: CE
authors.lastname: Barry
authors.forename: Honghai
authors.initials: H
authors.lastname: Wang
authors.forename: Wenhong
authors.initials: W
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000995:Antitubercular Agents / Q000378:metabolism / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D024881:Drug Resistance, Bacterial; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D011355:Prodrugs / Q000378:metabolism / Q000494:pharmacology; D011485:Protein Binding; D014176:Protein Biosynthesis / Q000187:drug effects; D017434:Protein Structure, Tertiary; D011718:Pyrazinamide / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology*; D012329:RNA, Bacterial / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D050505:Mutant Proteins; D011355:Prodrugs; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012269:Ribosomal Proteins; C015494:ribosomal protein S1; C096222:tmRNA; D011718:Pyrazinamide; C005296:pyrazinoic acid; D012343:RNA, Transfer; D000581:Amidohydrolases; C022490:pyrazinamide deamidase