VapCs of Mycobacterium tuberculosis cleave RNAs essential for translation.
The major human pathogen Mycobacterium tuberculosis can survive in the host organism for decades without causing symptoms. A large cohort of Toxin-Antitoxin (TA) modules contribute to this persistence. Of these, 48 TA modules belong to the vapBC (virulence associated protein) gene family. VapC toxins are PIN domain endonucleases that, in enterobacteria, inhibit translation by site-specific cleavage of initiator tRNA. In contrast, VapC20 of M. tuberculosis inhibits translation by site-specific cleavage of the universally conserved Sarcin-Ricin loop (SRL) in 23S rRNA. Here we identify the cellular targets of 12 VapCs from M. tuberculosis by applying UV-crosslinking and deep sequencing. Remarkably, these VapCs are all endoribonucleases that cleave RNAs essential for decoding at the ribosomal A-site. Eleven VapCs cleave specific tRNAs while one exhibits SRL cleavage activity. These findings suggest that multiple vapBC modules contribute to the survival of M. tuberculosis in its human host by reducing the level of translation.
Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, DK-2200 Copenhagen N, Denmark kristoffer.winther@bio.ku.dk.|Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, Richardson Road, NE2 4AX, Newcastle upon Tyne, UK.
Kristoffer
K
Winther
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney 2033, Australia.
Jai J
JJ
Tree
Wellcome Trust Centre for Cell Biology, University of Edinburgh, Michael Swann Building, King's Buildings, Edinburgh EH9 3BF, UK.
David
D
Tollervey
Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, DK-2200 Copenhagen N, Denmark kgerdes@bio.ku.dk.|Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, Richardson Road, NE2 4AX, Newcastle upon Tyne, UK.
Kenn
K
Gerdes
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D020459:Genes, rRNA; D054884:Host-Pathogen Interactions; D006801:Humans; D008958:Models, Molecular; D008968:Molecular Conformation; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D011485:Protein Binding; D014176:Protein Biosynthesis; D020871:RNA Stability; D012358:RNA, Transfer, Met / Q000737:chemistry / Q000378:metabolism*; D014376:Tuberculosis / Q000235:genetics* / Q000382:microbiology*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D012358:RNA, Transfer, Met; C000592554:VapC protein, Mycobacterium smegmatis