1vfg
From Proteopedia
(New page: 200px<br /><applet load="1vfg" size="450" color="white" frame="true" align="right" spinBox="true" caption="1vfg, resolution 2.8Å" /> '''Crystal structure of ...) |
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| - | [[Image:1vfg.gif|left|200px]]<br /><applet load="1vfg" size=" | + | [[Image:1vfg.gif|left|200px]]<br /><applet load="1vfg" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1vfg, resolution 2.8Å" /> | caption="1vfg, resolution 2.8Å" /> | ||
'''Crystal structure of tRNA nucleotidyltransferase complexed with a primer tRNA and an incoming ATP analog'''<br /> | '''Crystal structure of tRNA nucleotidyltransferase complexed with a primer tRNA and an incoming ATP analog'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The 3'-terminal CCA nucleotide sequence (positions 74-76) of transfer RNA | + | 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== | ==About this Structure== | ||
| - | 1VFG is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Aquifex_aeolicus Aquifex aeolicus] with APC as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Polynucleotide_adenylyltransferase Polynucleotide adenylyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.19 2.7.7.19] Full crystallographic information is available from [http:// | + | 1VFG is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Aquifex_aeolicus Aquifex aeolicus] with <scene name='pdbligand=APC:'>APC</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Polynucleotide_adenylyltransferase Polynucleotide adenylyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.19 2.7.7.19] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1VFG OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Ishitani, R.]] | [[Category: Ishitani, R.]] | ||
[[Category: Nureki, O.]] | [[Category: Nureki, O.]] | ||
| - | [[Category: RSGI, RIKEN | + | [[Category: RSGI, RIKEN Structural Genomics/Proteomics Initiative.]] |
[[Category: Takeuchi, N.]] | [[Category: Takeuchi, N.]] | ||
[[Category: Tomita, K.]] | [[Category: Tomita, K.]] | ||
[[Category: Ueda, T.]] | [[Category: Ueda, T.]] | ||
| - | [[Category: Vassylyev, D | + | [[Category: Vassylyev, D G.]] |
[[Category: APC]] | [[Category: APC]] | ||
[[Category: riken structural genomics/proteomics initiative]] | [[Category: riken structural genomics/proteomics initiative]] | ||
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[[Category: transferase]] | [[Category: transferase]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:34:42 2008'' |
Revision as of 13:34, 21 February 2008
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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 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 Thu Feb 21 15:34:42 2008
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
