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. 1998 Dec;180(24):6446-9.
doi: 10.1128/JB.180.24.6446-6449.1998.

Sequence divergence of seryl-tRNA synthetases in archaea

H S Kim  1 , U C Vothknecht, R Hedderich, I Celic, D Söll
Affiliations

Sequence divergence of seryl-tRNA synthetases in archaea

H S Kim et al. J Bacteriol. 1998 Dec.

Abstract

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.

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Figures

FIG. 1
FIG. 1
Northern blot analysis of unfractionated M. thermoautotrophicum tRNA charged with purified M. thermoautotrophicum SerRS. Charged and uncharged tRNA was separated (18), and the blots were probed with a tRNASer and tRNACys probe. Lanes: 1 and 3, uncharged tRNA; 2 and 5, tRNA charged with serine by M. thermoautotrophicum SerRS; 4, tRNA charged with cysteine by E. coli CysRS. AA, amino acid.
FIG. 2
FIG. 2
Alignment of motifs 1, 2, and 3 (4) from a number of representative SerRSs. The sequences (accession numbers) are from Homo sapiens (X91257) (Hsa), Saccharomyces cerevisiae (X04884) (Sce), Arabidopsis thaliana (Z70313) (Ath), Drosophila melanogaster (Y14823) (Dme), Escherichia coli (X04017) (Eco), Bacillus subtilis (D26185) (Bsu), Thermus aquaticus (sp:P34945) (Taq), Haloarcula marismortui (X91007) (Hma), Archaeoglobus fulgidus (AE000962) (Afu), Pyrococcus horikoshii (AB009490) (Pho), Methanococcus jannaschii (U67550) (Mja), Methanobacterium thermoautotrophicum (AF009823) (Mth), and Methanococcus maripaludis (AF009822) (Mma). They were aligned with the Clustal program (16), and the motif regions are presented.
FIG. 3
FIG. 3
Phylogenetic analysis of SerRS alignments by maximum likelihood analysis, using the Puzzle program (16).
See this image and copyright information in PMC

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