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1vfg
From Proteopedia
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Crystal structure of tRNA nucleotidyltransferase complexed with a primer tRNA and an incoming ATP analog
Overview
The 3'-terminal CCA nucleotide sequence (positions 74-76) of transfer RNA, is essential for amino acid attachment and interaction with the ribosome, during protein synthesis. The CCA sequence is synthesized de novo and/or, repaired by a template-independent RNA polymerase, 'CCA-adding enzyme', using CTP and ATP as substrates. Despite structural and biochemical, studies, the mechanism by which the CCA-adding enzyme synthesizes the, defined sequence without a nucleic acid template remains elusive. Here we, present the crystal structure of Aquifex aeolicus CCA-adding enzyme, bound, to a primer tRNA lacking the terminal adenosine and an incoming ATP, analogue, at 2.8 A resolution. The enzyme enfolds the acceptor T helix of, the tRNA molecule. In the catalytic pocket, C75 is adjacent to ATP, and, their base moieties are stacked. The complementary pocket for recognizing, C74-C75 of tRNA forms a 'protein template' for the penultimate two, nucleotides, mimicking the nucleotide template used by template-dependent, polymerases. These results are supported by systematic analyses of, mutants. Our structure represents the 'pre-insertion' stage of selecting, the incoming nucleotide and provides the structural basis for the, mechanism underlying template-independent RNA polymerization.
About this Structure
1VFG is a Single protein structure of sequence from Aquifex aeolicus with APC as ligand. Active as Polynucleotide adenylyltransferase, with EC number 2.7.7.19 Full crystallographic information is available from OCA.
Reference
Structural basis for template-independent RNA polymerization., Tomita K, Fukai S, Ishitani R, Ueda T, Takeuchi N, Vassylyev DG, Nureki O, Nature. 2004 Aug 5;430(7000):700-4. PMID:15295603
Page seeded by OCA on Sat Nov 24 22:20:14 2007
Categories: Aquifex aeolicus | Polynucleotide adenylyltransferase | Single protein | Fukai, S. | Ishitani, R. | Nureki, O. | RSGI, RIKEN.Structural.Genomics/Proteomics.Initiative. | Takeuchi, N. | Tomita, K. | Ueda, T. | Vassylyev, D.G. | APC | Riken structural genomics/proteomics initiative | Rna | Rsgi | Structural genomics | Transferase
