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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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