Abstract
AbstractTransfer (t)RNAs contain a wide variety of post-transcriptional modifications, which play critical roles in tRNA stability and functions. 3-(3-amino-3-carboxypropyl)uridine (acp3U) is a highly conserved modification found in variable- and D-loops of tRNAs. Biogenesis and functions of acp3U have not been extensively investigated. Using a reverse-genetic approach supported by comparative genomics, we find here that theEscherichia coli yfiPgene, which we renametapT(tRNA aminocarboxypropyltransferase), is responsible for acp3U formation in tRNA. Recombinant TapT synthesizes acp3U at position 47 of tRNAs in the presence ofS-adenosylmethionine. Biochemical experiments reveal that acp3U47 confers thermal stability on tRNA. Curiously, the ΔtapTstrain exhibits genome instability under continuous heat stress. We also find that the human homologs oftapT,DTWD1andDTWD2, are responsible for acp3U formation at positions 20 and 20a of tRNAs, respectively. Double knockout cells ofDTWD1andDTWD2exhibit growth retardation, indicating that acp3U is physiologically important in mammals.
Funder
MEXT | Japan Society for the Promotion of Science
Publisher
Springer Science and Business Media LLC
Subject
General Physics and Astronomy,General Biochemistry, Genetics and Molecular Biology,General Chemistry
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