2j6s: Difference between revisions
No edit summary |
No edit summary |
||
| Line 1: | Line 1: | ||
[[Image:2j6s.gif|left|200px]] | [[Image:2j6s.gif|left|200px]] | ||
<!-- | |||
The line below this paragraph, containing "STRUCTURE_2j6s", creates the "Structure Box" on the page. | |||
You may change the PDB parameter (which sets the PDB file loaded into the applet) | |||
or the SCENE parameter (which sets the initial scene displayed when the page is loaded), | |||
or leave the SCENE parameter empty for the default display. | |||
| | --> | ||
| | {{STRUCTURE_2j6s| PDB=2j6s | SCENE= }} | ||
}} | |||
'''TERNARY COMPLEX OF SULFOLOBUS SOLFATARICUS DPO4 DNA POLYMERASE, O6-METHYLGUANINE MODIFIED DNA, AND DATP.''' | '''TERNARY COMPLEX OF SULFOLOBUS SOLFATARICUS DPO4 DNA POLYMERASE, O6-METHYLGUANINE MODIFIED DNA, AND DATP.''' | ||
| Line 30: | Line 27: | ||
[[Category: Guengerich, F P.]] | [[Category: Guengerich, F P.]] | ||
[[Category: Irimia, A.]] | [[Category: Irimia, A.]] | ||
[[Category: | [[Category: Dna damage]] | ||
[[Category: | [[Category: Dna polymerase]] | ||
[[Category: | [[Category: Dna repair]] | ||
[[Category: | [[Category: Dna replication]] | ||
[[Category: | [[Category: Dna-binding]] | ||
[[Category: | [[Category: Dna-directed dna polymerase]] | ||
[[Category: | [[Category: Dpo4]] | ||
[[Category: | [[Category: Magnesium]] | ||
[[Category: | [[Category: Metal-binding]] | ||
[[Category: | [[Category: Mutator protein]] | ||
[[Category: | [[Category: Nucleotidyltransferase]] | ||
[[Category: | [[Category: O6-methylguanine]] | ||
[[Category: | [[Category: Sulfolobus solfataricus]] | ||
[[Category: | [[Category: Transferase]] | ||
[[Category: | [[Category: Transferase/dna complex]] | ||
[[Category: | [[Category: Translesion dna synthesis]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May 4 08:25:43 2008'' | |||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | |||
Revision as of 05:25, 4 May 2008
TERNARY COMPLEX OF SULFOLOBUS SOLFATARICUS DPO4 DNA POLYMERASE, O6-METHYLGUANINE MODIFIED DNA, AND DATP.
Overview
We examined the effect of a single O6-methylguanine (O6-MeG) template residue on catalysis by a model Y family polymerase, Dpo4 from Sulfolobus solfataricus. Mass spectral analysis of Dpo4-catalyzed extension products revealed that the enzyme accurately bypasses O6-MeG, with C being the major product (approximately 70%) and T or A being the minor species (approximately 20% or approximately 10%, respectively), consistent with steady-state kinetic parameters. Transient-state kinetic experiments revealed that kpol, the maximum forward rate constant describing polymerization, for dCTP incorporation opposite O6-MeG was approximately 6-fold slower than observed for unmodified G, and no measurable product was observed for dTTP incorporation in the pre-steady state. The lack of any structural information regarding how O6-MeG paired in a polymerase active site led us to perform x-ray crystallographic studies, which show that "wobble" pairing occurs between C and O6-MeG. A structure containing T opposite O6-MeG was solved, but much of the ribose and pyrimidine base density was disordered, in accordance with a much higher Km,dTTP that drives the difference in efficiency between C and T incorporation. The more stabilized C:O6-MeG pairing reinforces the importance of hydrogen bonding with respect to nucleotide selection within a geometrically tolerant polymerase active site.
About this Structure
2J6S is a Single protein structure of sequence from Sulfolobus solfataricus. Full crystallographic information is available from OCA.
Reference
Sulfolobus solfataricus DNA polymerase Dpo4 is partially inhibited by "wobble" pairing between O6-methylguanine and cytosine, but accurate bypass is preferred., Eoff RL, Irimia A, Egli M, Guengerich FP, J Biol Chem. 2007 Jan 12;282(2):1456-67. Epub 2006 Nov 14. PMID:17105728 Page seeded by OCA on Sun May 4 08:25:43 2008
Proteopedia Page Contributors and Editors (what is this?)
- Pages with broken file links
- DNA-directed DNA polymerase
- Single protein
- Sulfolobus solfataricus
- Egli, M.
- Eoff, R L.
- Guengerich, F P.
- Irimia, A.
- Dna damage
- Dna polymerase
- Dna repair
- Dna replication
- Dna-binding
- Dna-directed dna polymerase
- Dpo4
- Magnesium
- Metal-binding
- Mutator protein
- Nucleotidyltransferase
- O6-methylguanine
- Transferase
- Transferase/dna complex
- Translesion dna synthesis