Strand annealing and terminal transferase activities of a B-family DNA polymerase.
Abstract:
DNA replication polymerases have the inherent ability to faithfully and rapidly copy a DNA template according to precise Watson-Crick base pairing. The primary B-family DNA replication polymerase (Dpo1) in the hyperthermophilic archaeon, Sulfolobus solfataricus, is shown here to possess a remarkable DNA stabilizing ability for maintaining weak base pairing interactions to facilitate primer extension. This thermal stabilization by Dpo1 allowed for template-directed synthesis at temperatures more than 30 °C above the melting temperature of naked DNA. Surprisingly, Dpo1 also displays a competing terminal deoxynucleotide transferase (TdT) activity unlike any other B-family DNA polymerase. Dpo1 is shown to elongate single-stranded DNA in template-dependent and template-independent manners. Experiments with different homopolymeric templates indicate that initial deoxyribonucleotide incorporation is complementary to the template. Rate-limiting steps that include looping back and annealing to the template allow for a unique template-dependent terminal transferase activity. The multiple activities of this unique B-family DNA polymerase make this enzyme an essential component for DNA replication and DNA repair for the maintenance of the archaeal genome at high temperatures.
Polymerases:
Topics:
Historical Protein Properties (MW, pI, ...), Kinetic Parameters, Nucleotide Incorporation, Exonuclease Activity, Terminal Transferase, Source / Purification, Other
Status:
| new | topics/pols set | partial results | complete | validated |
Results:
| Polymerase | Reference | Property | Result | Context |
|---|---|---|---|---|
| SsoDpo1 | Strand annealing and terminal transferase activities of a B-family DNA polymerase. | Terminal Transferase | Yes | |
| SsoDpo1 | Strand annealing and terminal transferase activities of a B-family DNA polymerase. | Molecular Weight | 101kD | Technique: SDS-PAGE |
| SsoDpo1 | Strand annealing and terminal transferase activities of a B-family DNA polymerase. | 3-5' Exonuclease (proofreading) | Yes | |
| SsoDpo1 | Strand annealing and terminal transferase activities of a B-family DNA polymerase. | Cloned or native | Cloned in E. coli | |
| SsoDpo1 | Strand annealing and terminal transferase activities of a B-family DNA polymerase. | Tagged | No | |
| SsoDpo1 | Strand annealing and terminal transferase activities of a B-family DNA polymerase. | Full length or truncated | Full length | |
| SsoDpo1 | Strand annealing and terminal transferase activities of a B-family DNA polymerase. | Maximum Product Length | 10kb |