Structural basis of RNA-dependent recruitment of glutamine to the genetic code

Glutaminyl-transfer RNA (Gln-tRNA(Gln)) in archaea is synthesized in a pretranslational amidation of misacylated Glu-tRNA(Gln) by the heterodimeric Glu-tRNA(Gln) amidotransferase GatDE. Here we report the crystal structure of the Methanothermobacter thermautotrophicus GatDE complexed to tRNA(Gln) at...

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Published in:Science (American Association for the Advancement of Science) Vol. 312; no. 5782; p. 1950
Main Authors: Oshikane, Hiroyuki, Sheppard, Kelly, Fukai, Shuya, Nakamura, Yuko, Ishitani, Ryuichiro, Numata, Tomoyuki, Sherrer, R Lynn, Feng, Liang, Schmitt, Emmanuelle, Panvert, Michel, Blanquet, Sylvain, Mechulam, Yves, Söll, Dieter, Nureki, Osamu
Format: Journal Article
Language:English
Published: United States 30.06.2006
Subjects:
Acylation
Adenosine Triphosphate - metabolism
Ammonia - metabolism
Anticodon
Binding Sites
Catalytic Domain
Computer Simulation
Crystallography, X-Ray
Dimerization
Genetic Code
Glutamine - metabolism
Hydrogen Bonding
Magnesium - metabolism
Methanobacteriaceae - enzymology
Methanobacteriaceae - genetics
Models, Molecular
Mutation
Nitrogenous Group Transferases - chemistry
Nitrogenous Group Transferases - metabolism
Nucleic Acid Conformation
Protein Structure, Quaternary
Protein Structure, Secondary
Protein Structure, Tertiary
RNA, Archaeal - chemistry
RNA, Archaeal - metabolism
RNA, Transfer, Gln - chemistry
RNA, Transfer, Gln - metabolism
ISSN:1095-9203, 1095-9203
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Summary:Glutaminyl-transfer RNA (Gln-tRNA(Gln)) in archaea is synthesized in a pretranslational amidation of misacylated Glu-tRNA(Gln) by the heterodimeric Glu-tRNA(Gln) amidotransferase GatDE. Here we report the crystal structure of the Methanothermobacter thermautotrophicus GatDE complexed to tRNA(Gln) at 3.15 angstroms resolution. Biochemical analysis of GatDE and of tRNA(Gln) mutants characterized the catalytic centers for the enzyme's three reactions (glutaminase, kinase, and amidotransferase activity). A 40 angstrom-long channel for ammonia transport connects the active sites in GatD and GatE. tRNA(Gln) recognition by indirect readout based on shape complementarity of the D loop suggests an early anticodon-independent RNA-based mechanism for adding glutamine to the genetic code.
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ISSN:1095-9203
1095-9203
DOI:10.1126/science.1128470