1: Nucleic Acids Res. 1990 Dec 11;18(23):7109-18.

Aspartyl-tRNA synthetase from Escherichia coli: cloning and characterisation of
the gene, homologies of its translated amino acid sequence with asparaginyl- and 
lysyl-tRNA synthetases.

Eriani G, Dirheimer G, Gangloff J.

Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France.

By screening of an Escherichia coli plasmidic library using antibodies against
aspartyl-tRNA synthetase (AspRS) several clones were obtained containing aspS,
the gene coding for AspRS. We report here the nucleotide sequence of aspS and the
corresponding primary structure of the aspartyl-tRNA synthetase, a protein of 590
amino acid residues with a Mr 65,913, a value in close agreement with that
observed for the purified protein. Primer extension analysis of the aspS mRNA
using reverse transcriptase located its 5'-end at 94 nucleotides upstream of the 
translation initiation AUG; nuclease S1 analysis located the 3'-end at 126
nucleotides downstream of the stop codon UGA. Comparison of the DNA-derived
protein sequence with known aminoacyl-tRNA sequences revealed important
homologies with asparaginyl- and lysyl-tRNA synthetases from E.coli; more than
25% of their amino acid residues are identical, the homologies being distributed 
preferencially in the first part and the carboxy-terminal end of the molecule.
Mutagenesis directed towards a consensus tetrapeptide (Gly-Leu-Asp-Arg) and the
carboxy-terminal end showed that both domains could be implicated in catalysis as
well as in ATP binding.

Publication Types: 
    Research Support, Non-U.S. Gov't

PMID: 2129559 [PubMed - indexed for MEDLINE]