Hybrid Methods Reveal Multiple Flexibly Linked DNA Polymerases within the Bacteriophage T7 Replisome
Abstract
The physical organization of DNA enzymes at a replication fork enables efficient copying of two antiparallel DNA strands, yet dynamic protein interactions within the replication complex complicate replisome structural studies. We employed a combination of crystallographic, native mass spectrometry and small-angle X-ray scattering experiments to capture alternative structures of a model replication system encoded by bacteriophage T7. then, the two molecules of DNA polymerase bind the ring-shaped primase-helicase in a conserved orientation and provide structural insight into how the acidic C-terminal tail of the primase-helicase contacts the DNA polymerase to facilitate loading of the polymerase onto DNA. A third DNA polymerase binds the ring in an offset manner that may enable polymerase exchange during replication. Alternative polymerase binding modes are also detected by small-angle X-ray scattering with DNA substrates present. The collective results unveil complex motions within T7 replisome higher-order structures that are underpinned by multivalent protein-protein interactions with functional implications.
- Authors:
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC); National Institutes of Health (NIH)
- OSTI Identifier:
- 1415494
- Alternate Identifier(s):
- OSTI ID: 1397618; OSTI ID: 1415964
- Grant/Contract Number:
- SC0001035; AC02-05CH11231
- Resource Type:
- Published Article
- Journal Name:
- Structure
- Additional Journal Information:
- Journal Name: Structure Journal Volume: 25 Journal Issue: 1; Journal ID: ISSN 0969-2126
- Publisher:
- Elsevier
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; DNA replication; x-ray crystallography; native mass spectrometry; small-angle x-ray scattering
Citation Formats
Wallen, Jamie R., Zhang, Hao, Weis, Caroline, Cui, Weidong, Foster, Brittni M., Ho, Chris M. W., Hammel, Michal, Tainer, John A., Gross, Michael L., and Ellenberger, Tom. Hybrid Methods Reveal Multiple Flexibly Linked DNA Polymerases within the Bacteriophage T7 Replisome. United Kingdom: N. p., 2017.
Web. doi:10.1016/j.str.2016.11.019.
Wallen, Jamie R., Zhang, Hao, Weis, Caroline, Cui, Weidong, Foster, Brittni M., Ho, Chris M. W., Hammel, Michal, Tainer, John A., Gross, Michael L., & Ellenberger, Tom. Hybrid Methods Reveal Multiple Flexibly Linked DNA Polymerases within the Bacteriophage T7 Replisome. United Kingdom. https://doi.org/10.1016/j.str.2016.11.019
Wallen, Jamie R., Zhang, Hao, Weis, Caroline, Cui, Weidong, Foster, Brittni M., Ho, Chris M. W., Hammel, Michal, Tainer, John A., Gross, Michael L., and Ellenberger, Tom. Sun .
"Hybrid Methods Reveal Multiple Flexibly Linked DNA Polymerases within the Bacteriophage T7 Replisome". United Kingdom. https://doi.org/10.1016/j.str.2016.11.019.
@article{osti_1415494,
title = {Hybrid Methods Reveal Multiple Flexibly Linked DNA Polymerases within the Bacteriophage T7 Replisome},
author = {Wallen, Jamie R. and Zhang, Hao and Weis, Caroline and Cui, Weidong and Foster, Brittni M. and Ho, Chris M. W. and Hammel, Michal and Tainer, John A. and Gross, Michael L. and Ellenberger, Tom},
abstractNote = {The physical organization of DNA enzymes at a replication fork enables efficient copying of two antiparallel DNA strands, yet dynamic protein interactions within the replication complex complicate replisome structural studies. We employed a combination of crystallographic, native mass spectrometry and small-angle X-ray scattering experiments to capture alternative structures of a model replication system encoded by bacteriophage T7. then, the two molecules of DNA polymerase bind the ring-shaped primase-helicase in a conserved orientation and provide structural insight into how the acidic C-terminal tail of the primase-helicase contacts the DNA polymerase to facilitate loading of the polymerase onto DNA. A third DNA polymerase binds the ring in an offset manner that may enable polymerase exchange during replication. Alternative polymerase binding modes are also detected by small-angle X-ray scattering with DNA substrates present. The collective results unveil complex motions within T7 replisome higher-order structures that are underpinned by multivalent protein-protein interactions with functional implications.},
doi = {10.1016/j.str.2016.11.019},
journal = {Structure},
number = 1,
volume = 25,
place = {United Kingdom},
year = {Sun Jan 01 00:00:00 EST 2017},
month = {Sun Jan 01 00:00:00 EST 2017}
}
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https://doi.org/10.1016/j.str.2016.11.019
https://doi.org/10.1016/j.str.2016.11.019
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Cited by: 14 works
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Works referencing / citing this record:
Cryo-EM structure of the replisome reveals multiple interactions coordinating DNA synthesis
journal, February 2017
- Kulczyk, Arkadiusz W.; Moeller, Arne; Meyer, Peter
- Proceedings of the National Academy of Sciences, Vol. 114, Issue 10
Structural basis for adPEO-causing mutations in the mitochondrial TWINKLE helicase
journal, November 2018
- Peter, Bradley; Farge, Geraldine; Pardo-Hernandez, Carlos
- Human Molecular Genetics, Vol. 28, Issue 7
Helicase promotes replication re-initiation from an RNA transcript
journal, June 2018
- Sun, Bo; Singh, Anupam; Sultana, Shemaila
- Nature Communications, Vol. 9, Issue 1
The ring-shaped hexameric helicases that function at DNA replication forks
journal, January 2018
- O’Donnell, Michael E.; Li, Huilin
- Nature Structural & Molecular Biology, Vol. 25, Issue 2
Structures and operating principles of the replisome
journal, January 2019
- Gao, Yang; Cui, Yanxiang; Fox, Tara
- Science, Vol. 363, Issue 6429