Essentiality of threonylcarbamoyladenosine (t(6)A), a universal tRNA modification, in bacteria.
Threonylcarbamoyladenosine (t(6)A) is a modified nucleoside universally conserved in tRNAs in all three kingdoms of life. The recently discovered genes for t(6)A synthesis, including tsaC and tsaD, are essential in model prokaryotes but not essential in yeast. These genes had been identified as antibacterial targets even before their functions were known. However, the molecular basis for this prokaryotic-specific essentiality has remained a mystery. Here, we show that t(6)A is a strong positive determinant for aminoacylation of tRNA by bacterial-type but not by eukaryotic-type isoleucyl-tRNA synthetases and might also be a determinant for the essential enzyme tRNA(Ile)-lysidine synthetase. We confirm that t(6)A is essential in Escherichia coli and a survey of genome-wide essentiality studies shows that genes for t(6)A synthesis are essential in most prokaryotes. This essentiality phenotype is not universal in Bacteria as t(6)A is dispensable in Deinococcus radiodurans, Thermus thermophilus, Synechocystis PCC6803 and Streptococcus mutans. Proteomic analysis of t(6)A(-) D. radiodurans strains revealed an induction of the proteotoxic stress response and identified genes whose translation is most affected by the absence of t(6)A in tRNAs. Thus, although t(6)A is universally conserved in tRNAs, its role in translation might vary greatly between organisms.
Department of Microbiology and Cell Science, University of Florida, Gainesville, FL, 32611, USA.|Genetics and Genomics Graduate Program, University of Florida, Gainesville, FL, 32610, USA.|University of Florida Genetics Institute, University of Florida, Gainesville, FL, 32610, USA.|Institut de Génétique et Microbiologie, Université of Paris-Sud, Orsay, France.
Patrick C
PC
Thiaville
Department of Microbiology and Cell Science, University of Florida, Gainesville, FL, 32611, USA.
Basma
B
El Yacoubi
Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Caroline
C
Köhrer
Department of Microbiology and Cell Science, University of Florida, Gainesville, FL, 32611, USA.
Jennifer J
JJ
Thiaville
Department of Chemistry, Portland State University, Portland, OR, 97297, USA.
Chris
C
Deutsch
Department of Chemistry, Portland State University, Portland, OR, 97297, USA.
Dirk
D
Iwata-Reuyl
Department of Microbiology and Cell Science, University of Florida, Gainesville, FL, 32611, USA.
Jo Marie
JM
Bacusmo
CEA, DSV, IBiTec-S, SPI, Li2D, Laboratory 'Innovative technologies for Detection and Diagnostics', Bagnols-sur-Cèze, F-30200, France.
Jean
J
Armengaud
RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo, Hyogo, 679-5148, Japan.|Institute of Physics, Academia Sinica, 128 Sec. 2, Academia Rd., Nankang, Taipei, 11529, Taiwan.
Yoshitaka
Y
Bessho
Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, Cincinnati, Ohio, 45221, USA.
Collin
C
Wetzel
Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, Cincinnati, Ohio, 45221, USA.
Xiaoyu
X
Cao
Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, Cincinnati, Ohio, 45221, USA.
Patrick A
PA
Limbach
Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Uttam L
UL
RajBhandary
Department of Microbiology and Cell Science, University of Florida, Gainesville, FL, 32611, USA.|University of Florida Genetics Institute, University of Florida, Gainesville, FL, 32610, USA.
Valérie
V
de Crécy-Lagard
D000241:Adenosine / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics / Q000378:metabolism; D049148:Aminoacylation / Q000235:genetics; D017124:Conserved Sequence; D034301:Deinococcus / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011387:Prokaryotic Cells; D040901:Proteomics; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism; D012343:RNA, Transfer / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics
D012329:RNA, Bacterial; C003268:N(6)-(N-threonylcarbonyl)adenosine; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases; D000241:Adenosine