+Search query
-Structure paper
Title | Identification and characterization of a heterotrimeric archaeal DNA polymerase holoenzyme. |
---|---|
Journal, issue, pages | Nat Commun, Vol. 8, Page 15075, Year 2017 |
Publish date | May 2, 2017 |
Authors | Jiangyu Yan / Thomas R Beattie / Adriana L Rojas / Kelly Schermerhorn / Tamzin Gristwood / Jonathan C Trinidad / Sonja V Albers / Pietro Roversi / Andrew F Gardner / Nicola G A Abrescia / Stephen D Bell / |
PubMed Abstract | Since their initial characterization over 30 years ago, it has been believed that the archaeal B-family DNA polymerases are single-subunit enzymes. This contrasts with the multi-subunit B-family ...Since their initial characterization over 30 years ago, it has been believed that the archaeal B-family DNA polymerases are single-subunit enzymes. This contrasts with the multi-subunit B-family replicative polymerases of eukaryotes. Here we reveal that the highly studied PolB1 from Sulfolobus solfataricus exists as a heterotrimeric complex in cell extracts. Two small subunits, PBP1 and PBP2, associate with distinct surfaces of the larger catalytic subunit and influence the enzymatic properties of the DNA polymerase. Thus, multi-subunit replicative DNA polymerase holoenzymes are present in all three domains of life. We reveal the architecture of the assembly by a combination of cross-linking coupled with mass spectrometry, X-ray crystallography and single-particle electron microscopy. The small subunits stabilize the holoenzyme assembly and the acidic tail of one small subunit mitigates the ability of the enzyme to perform strand-displacement synthesis, with important implications for lagging strand DNA synthesis. |
External links | Nat Commun / PubMed:28462924 / PubMed Central |
Methods | EM (single particle) / X-ray diffraction |
Resolution | 1.351 - 22.8 Å |
Structure data | EMDB-3458: EMDB-3462: PDB-5n35: PDB-5n41: |
Chemicals | ChemComp-GD: ChemComp-GOL: ChemComp-NO3: ChemComp-HOH: ChemComp-EDO: |
Source |
|
Keywords | POLYMERASE BINDING PROTEIN / PolB1 binding protein 2 / Archaeal DNA Polymerase Holoenzyme / Protein binding / Polymerase-binding protein |