Please use this identifier to cite or link to this item: http://hdl.handle.net/2381/43088
Title: Identification and characterization of a heterotrimeric archaeal DNA polymerase holoenzyme.
Authors: Yan, Jiangyu
Beattie, Thomas R.
Rojas, Adriana L.
Schermerhorn, Kelly
Gristwood, Tamzin
Trinidad, Jonathan C.
Albers, Sonja V.
Roversi, Pietro
Gardner, Andrew F.
Abrescia, Nicola G. A.
Bell, Stephen D.
First Published: 2-May-2017
Publisher: Nature Research (part of Springer Nature)
Citation: Nature Communications, 2017, 8, 15075
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 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.
DOI Link: 10.1038/ncomms15075
eISSN: 2041-1723
Links: https://www.nature.com/articles/ncomms15075
http://hdl.handle.net/2381/43088
Version: Publisher Version
Status: Peer-reviewed
Type: Journal Article
Rights: Copyright © the authors, 2017. This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Description: Supplementary Information accompanies this paper at http://www.nature.com/ naturecommunications Protein Data Bank: Atomic coordinates and structure factors have been deposited under the accession number 5N35 (Gd-PBP2) and 5N41 (PBP2). Electron Microscopy Bank: Maps corresponding to the Apo and holo structures have been deposited with the accession numbers EMD-3458 (apo-PolB1), EMD3462 (PolB1-HE).
Appears in Collections:Published Articles, Dept. of Molecular and Cell Biology

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