Crystal structure of a thermostable type B DNA polymerase from Thermococcus gorgonarius.

Abstract:

Most known archaeal DNA polymerases belong to the type B family, which also includes the DNA replication polymerases of eukaryotes, but maintain high fidelity at extreme conditions. We describe here the 2.5 A resolution crystal structure of a DNA polymerase from the Archaea Thermococcus gorgonarius and identify structural features of the fold and the active site that are likely responsible for its thermostable function. Comparison with the mesophilic B type DNA polymerase gp43 of the bacteriophage RB69 highlights thermophilic adaptations, which include the presence of two disulfide bonds and an enhanced electrostatic complementarity at the DNA-protein interface. In contrast to gp43, several loops in the exonuclease and thumb domains are more closely packed; this apparently blocks primer binding to the exonuclease active site. A physiological role of this "closed" conformation is unknown but may represent a polymerase mode, in contrast to an editing mode with an open exonuclease site. This archaeal B DNA polymerase structure provides a starting point for structure-based design of polymerases or ligands with applications in biotechnology and the development of antiviral or anticancer agents.

Polymerases:

TGO

Topics:

Structure and Structure/Function, Exonuclease Activity

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
TGO Hopfner KP1999 3-5' Exonuclease (proofreading) Yes
TGO Hopfner KP1999 Cloned or native Cloned in yeast
TGO Hopfner KP1999 Amino Acids Contacting Primer Tyr402
TGO Hopfner KP1999 Full length or truncated Full length
TGO Hopfner KP1999 Amino Acids Contacting NTP K487x3NSxYGx2G

Entry validated by:

Structures:

1TGO
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