title: Sequence divergence of seryl-tRNA synthetases in archaea.
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.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Cellular and Developmental Biology, Yale University, New Haven, Connecticut 06520-8114, USA.
authors.forename: H S
authors.initials: HS
authors.lastname: Kim
authors.forename: U C
authors.initials: UC
authors.lastname: Vothknecht
authors.forename: R
authors.initials: R
authors.lastname: Hedderich
authors.forename: I
authors.initials: I
authors.lastname: Celic
authors.forename: D
authors.initials: D
authors.lastname: Söll
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D019641:DNA, Archaeal; D015870:Gene Expression; D006801:Humans; D017014:Methanobacterium / Q000201:enzymology* / Q000235:genetics; D017017:Methanococcus / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D019642:RNA, Archaeal / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012714:Serine-tRNA Ligase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D019641:DNA, Archaeal; D019642:RNA, Archaeal; D012343:RNA, Transfer; D012714:Serine-tRNA Ligase