Human cytoplasmic isoleucyl-tRNA synthetase: selective divergence of the anticodon-binding domain and acquisition of a new structural unit.
We show here that the class I human cytoplasmic isoleucyl-tRNA synthetase is an exceptionally large polypeptide (1266 aa) which, unlike its homologues in lower eukaryotes and prokaryotes, has a third domain of two repeats of an approximately 90-aa sequence appended to its C-terminal end. While extracts of Escherichia coli do not aminoacrylate mammalian tRNA with isoleucine, expression of the cloned human gene in E. coli results in charging of the mammalian tRNA substrate. The appended third domain is dispensable for detection of this aminoacylation activity and may be needed for assembly of a multisynthetase complex in mammalian cells. Alignment of the sequences of the remaining two domains shared by isoleucyl-tRNA synthetases from E. coli to human reveals a much greater selective pressure on the domain needed for tRNA acceptor helix interactions and catalysis than on the domain needed for interactions with the anticodon. This result may have implications for the historical development of an operational RNA code for amino acids.
Shiba
K
K
Department of Cell Biology, Cancer Institute, Tokyo, Japan.
Suzuki
N
N
Shigesada
K
K
Namba
Y
Y
Schimmel
P
P
Noda
T
T
D000595:Amino Acid Sequence; D002451:Cell Compartmentation; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000201:enzymology; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007533:Isoleucine-tRNA Ligase / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D012356:RNA, Transfer, Leu / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013329:Structure-Activity Relationship
D004251:DNA Transposable Elements; D012356:RNA, Transfer, Leu; D011994:Recombinant Proteins; D007533:Isoleucine-tRNA Ligase