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Journal of Bacteriology, December 1998, p. 6446-6449, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Sequence Divergence of Seryl-tRNA Synthetases
in Archaea
Hyun-soo
Kim,1
Ute C.
Vothknecht,1
Reiner
Hedderich,2
Ivana
Celic,1 and
Dieter
Söll1,3,*
Department of Molecular Biophysics and
Biochemistry1 and
Department of
Molecular, Cellular and Developmental
Biology,3 Yale University, New Haven,
Connecticut 06520-8114, and
Abteilung Biochemie,
Max-Planck-Institut für Terrestrische Mikrobiologie, D-35043
Marburg, Germany2
Received 26 June 1998/Accepted 28 September 1998
The genomic sequences of Methanococcus jannaschii and
Methanobacterium thermoautotrophicum contain a structurally
uncommon seryl-tRNA synthetase (SerRS) sequence and lack an open
reading frame (ORF) for the canonical cysteinyl-tRNA synthetase
(CysRS). Therefore, it is not clear if Cys-tRNACys is
formed by direct aminoacylation or by a transformation of serine
misacylated to tRNACys. To address this question, we
prepared SerRS from two methanogenic archaea and measured the enzymatic
properties of these proteins. SerRS was purified from M. thermoautotrophicum; its N-terminal peptide sequence matched the
sequence deduced from the relevant ORF in the genomic data of M. thermoautotrophicum and M. jannaschii. In addition,
SerRS was expressed from a cloned Methanococcus maripaludis serS gene. The two enzymes charged serine to their homologous tRNAs and also accepted Escherichia coli tRNA as substrate
for aminoacylation. Gel shift experiments showed that M. thermoautotrophicum SerRS did not mischarge tRNACys
with serine. This indicates that Cys-tRNACys is formed by
direct acylation in these organisms.
*
Corresponding author. Mailing address: Department of
Molecular Biophysics and Biochemistry, Yale University, P.O. Box
208114, 266 Whitney Ave., New Haven, CT 06520-8114. Phone: (203)
432-6200. Fax: (203) 432-6202. E-mail:
soll{at}trna.chem.yale.edu.
Journal of Bacteriology, December 1998, p. 6446-6449, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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