RRC ID 35411
Author Winther KS, Gerdes K.
Title Enteric virulence associated protein VapC inhibits translation by cleavage of initiator tRNA.
Journal Proc Natl Acad Sci U S A
Abstract Eukaryotic PIN (PilT N-terminal) domain proteins are ribonucleases involved in quality control, metabolism and maturation of mRNA and rRNA. The majority of prokaryotic PIN-domain proteins are encoded by the abundant vapBC toxin--antitoxin loci and inhibit translation by an unknown mechanism. Here we show that enteric VapCs are site-specific endonucleases that cleave tRNA(fMet) in the anticodon stem-loop between nucleotides +38 and +39 in vivo and in vitro. Consistently, VapC inhibited translation in vivo and in vitro. Translation-reactions could be reactivated by the addition of VapB and extra charged tRNA(fMet). Similarly, ectopic production of tRNA(fMet) counteracted VapC in vivo. Thus, tRNA(fMet) is the only cellular target of VapC. Depletion of tRNA(fMet) by vapC induction was bacteriostatic and stimulated ectopic translation initiation at elongator codons. Moreover, addition of chloramphenicol to cells carrying vapBC induced VapC activity. Thus, by cleavage of tRNA(fMet), VapC simultaneously may regulate global cellular translation and reprogram translation initiation.
Volume 108(18)
Pages 7403-7
Published 2011-5-3
DOI 10.1073/pnas.1019587108
PII 1019587108
PMID 21502523
PMC PMC3088637
MeSH Bacterial Proteins / metabolism* Blotting, Northern Blotting, Western Chloramphenicol Endoribonucleases / metabolism* Gene Expression Regulation, Bacterial / genetics Gene Expression Regulation, Bacterial / physiology* Luciferases Membrane Glycoproteins / metabolism* Oligonucleotides / genetics Plasmids / genetics RNA, Transfer, Met / metabolism* Salmonella typhimurium / enzymology* Shigella flexneri / enzymology* Ultracentrifugation
IF 9.412
Times Cited 159
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Prokaryotes E. coli ?