Transfer RNA-dependent amino acid biosynthesis: an essential route to asparagine formation.
Biochemical experiments and genomic sequence analysis showed that Deinococcus radiodurans and Thermus thermophilus do not possess asparagine synthetase (encoded by asnA or asnB), the enzyme forming asparagine from aspartate. Instead these organisms derive asparagine from asparaginyl-tRNA, which is made from aspartate in the tRNA-dependent transamidation pathway [Becker, H. D. & Kern, D. (1998) Proc. Natl. Acad. Sci. USA 95, 12832-12837; and Curnow, A. W., Tumbula, D. L., Pelaschier, J. T., Min, B. & Söll, D. (1998) Proc. Natl. Acad. Sci. USA 95, 12838-12843]. A genetic knockout disrupting this pathway deprives D. radiodurans of the ability to synthesize asparagine and confers asparagine auxotrophy. The organism's capacity to make asparagine could be restored by transformation with Escherichia coli asnB. This result demonstrates that in Deinococcus, the only route to asparagine is via asparaginyl-tRNA. Analysis of the completed genomes of many bacteria reveal that, barring the existence of an unknown pathway of asparagine biosynthesis, a wide spectrum of bacteria rely on the tRNA-dependent transamidation pathway as the sole route to asparagine.
Min
Bokkee
B
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520-8114, USA.
Pelaschier
Joanne T
JT
Graham
David E
DE
Tumbula-Hansen
Debra
D
Söll
Dieter
D
D000596:Amino Acids / Q000096:biosynthesis; D001216:Asparagine / Q000096:biosynthesis*; D001226:Aspartate-tRNA Ligase / Q000378:metabolism*; D001224:Aspartic Acid / Q000378:metabolism; D019733:Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005051:Eubacterium / Q000378:metabolism*; D012346:RNA, Transfer, Amino Acyl / Q000096:biosynthesis*
D000596:Amino Acids; D012346:RNA, Transfer, Amino Acyl; D001224:Aspartic Acid; D001216:Asparagine; D001226:Aspartate-tRNA Ligase; C029194:asparaginyl-tRNA synthetase; D019733:Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor; C019742:asparagine synthetase (glutamine-hydrolyzing)