The Bacillus subtilis tyrZ gene encodes a highly selective tyrosyl-tRNA synthetase and is regulated by a MarR regulator and T box riboswitch.
Misincorporation of D-tyrosine (D-Tyr) into cellular proteins due to mischarging of tRNA(Tyr) with D-Tyr by tyrosyl-tRNA synthetase inhibits growth and biofilm formation of Bacillus subtilis. Furthermore, many B. subtilis strains lack a functional gene encoding D-aminoacyl-tRNA deacylase, which prevents misincorporation of D-Tyr in most organisms. B. subtilis has two genes that encode tyrosyl-tRNA synthetase: tyrS is expressed under normal growth conditions, and tyrZ is known to be expressed only when tyrS is inactivated by mutation. We hypothesized that tyrZ encodes an alternate tyrosyl-tRNA synthetase, expression of which allows the cell to grow when D-Tyr is present. We show that TyrZ is more selective for L-Tyr over D-Tyr than is TyrS; however, TyrZ is less efficient overall. We also show that expression of tyrZ is required for growth and biofilm formation in the presence of D-Tyr. Both tyrS and tyrZ are preceded by a T box riboswitch, but tyrZ is found in an operon with ywaE, which is predicted to encode a MarR family transcriptional regulator. Expression of tyrZ is repressed by YwaE and also is regulated at the level of transcription attenuation by the T box riboswitch. We conclude that expression of tyrZ may allow growth when excess D-Tyr is present.


OBJECTIVE
Accurate protein synthesis requires correct aminoacylation of each tRNA with the cognate amino acid and discrimination against related compounds. Bacillus subtilis produces D-Tyr, an analog of L-Tyr that is toxic when incorporated into protein, during stationary phase. Most organisms utilize a D-aminoacyl-tRNA deacylase to prevent misincorporation of D-Tyr. This work demonstrates that the increased selectivity of the TyrZ form of tyrosyl-tRNA synthetase may provide a mechanism by which B. subtilis prevents misincorporation of D-Tyr in the absence of a functional D-aminoacyl-tRNA deacylase gene.
Department of Microbiology and Center for RNA Biology, The Ohio State University, Columbus, Ohio, USA.
Rebecca N
RN
Williams-Wagner
Department of Microbiology and Center for RNA Biology, The Ohio State University, Columbus, Ohio, USA.
Frank J
FJ
Grundy
Department of Microbiology and Center for RNA Biology, The Ohio State University, Columbus, Ohio, USA Ohio State Biochemistry Program, The Ohio State University, Columbus, Ohio, USA.
Medha
M
Raina
Department of Microbiology and Center for RNA Biology, The Ohio State University, Columbus, Ohio, USA Ohio State Biochemistry Program, The Ohio State University, Columbus, Ohio, USA.
Michael
M
Ibba
Department of Microbiology and Center for RNA Biology, The Ohio State University, Columbus, Ohio, USA Ohio State Biochemistry Program, The Ohio State University, Columbus, Ohio, USA henkin.3@osu.edu.
Tina M
TM
Henkin
D001412:Bacillus subtilis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D058928:Riboswitch; D013379:Substrate Specificity; D014157:Transcription Factors / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism; D014448:Tyrosine-tRNA Ligase / Q000235:genetics* / Q000378:metabolism*
D058928:Riboswitch; D014157:Transcription Factors; D014443:Tyrosine; D014448:Tyrosine-tRNA Ligase