2ago: Difference between revisions
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[[Image:2ago.gif|left|200px]] | [[Image:2ago.gif|left|200px]] | ||
'''Fidelity of Dpo4: effect of metal ions, nucleotide selection and pyrophosphorolysis''' | {{Structure | ||
|PDB= 2ago |SIZE=350|CAPTION= <scene name='initialview01'>2ago</scene>, resolution 2.850Å | |||
|SITE= | |||
|LIGAND= <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene> and <scene name='pdbligand=POP:PYROPHOSPHATE 2-'>POP</scene> | |||
|ACTIVITY= [http://en.wikipedia.org/wiki/DNA-directed_DNA_polymerase DNA-directed DNA polymerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.7 2.7.7.7] | |||
|GENE= dbh, dpo4 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=2287 Sulfolobus solfataricus]) | |||
}} | |||
'''Fidelity of Dpo4: effect of metal ions, nucleotide selection and pyrophosphorolysis''' | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
2AGO is a [ | 2AGO is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Sulfolobus_solfataricus Sulfolobus solfataricus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2AGO OCA]. | ||
==Reference== | ==Reference== | ||
Fidelity of Dpo4: effect of metal ions, nucleotide selection and pyrophosphorolysis., Vaisman A, Ling H, Woodgate R, Yang W, EMBO J. 2005 Sep 7;24(17):2957-67. Epub 2005 Aug 18. PMID:[http:// | Fidelity of Dpo4: effect of metal ions, nucleotide selection and pyrophosphorolysis., Vaisman A, Ling H, Woodgate R, Yang W, EMBO J. 2005 Sep 7;24(17):2957-67. Epub 2005 Aug 18. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16107880 16107880] | ||
[[Category: DNA-directed DNA polymerase]] | [[Category: DNA-directed DNA polymerase]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
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[[Category: pyrophosphorolysis]] | [[Category: pyrophosphorolysis]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 15:49:27 2008'' | ||
Revision as of 16:49, 20 March 2008
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| , resolution 2.850Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | , and | ||||||
| Gene: | dbh, dpo4 (Sulfolobus solfataricus) | ||||||
| Activity: | DNA-directed DNA polymerase, with EC number 2.7.7.7 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
Fidelity of Dpo4: effect of metal ions, nucleotide selection and pyrophosphorolysis
OverviewOverview
We report the crystal structures of a translesion DNA polymerase, Dpo4, complexed with a matched or mismatched incoming nucleotide and with a pyrophosphate product after misincorporation. These structures suggest two mechanisms by which Dpo4 may reject a wrong incoming nucleotide with its preformed and open active site. First, a mismatched replicating base pair leads to poor base stacking and alignment of the metal ions and as a consequence, inhibits incorporation. By replacing Mg2+ with Mn2+, which has a relaxed coordination requirement and tolerates misalignment, the catalytic efficiency of misincorporation increases dramatically. Mn2+ also enhances translesion synthesis by Dpo4. Subtle conformational changes that lead to the proper metal ion coordination may, therefore, be a key step in catalysis. Second, the slow release of pyrophosphate may increase the fidelity of Dpo4 by stalling mispaired primer extension and promoting pyrophosphorolysis that reverses the polymerization reaction. Indeed, Dpo4 has robust pyrophosphorolysis activity and degrades the primer strand in the presence of pyrophosphate. The correct incoming nucleotide allows DNA synthesis to overcome pyrophosphorolysis, but an incorrect incoming nucleotide does not.
About this StructureAbout this Structure
2AGO is a Single protein structure of sequence from Sulfolobus solfataricus. Full crystallographic information is available from OCA.
ReferenceReference
Fidelity of Dpo4: effect of metal ions, nucleotide selection and pyrophosphorolysis., Vaisman A, Ling H, Woodgate R, Yang W, EMBO J. 2005 Sep 7;24(17):2957-67. Epub 2005 Aug 18. PMID:16107880
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