DOI: 10.1073/pnas.93.23.12896. Reconstitution of human replication factor C from its five subunits in baculovirus-infected insect cells

Reconstitution of human replication factor C from its five subunits in baculovirus-infected insect cells

10.1073/pnas.93.23.12896

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Human replication factor C (RFC, also called activator 1) is a five-subunit protein complex (p140, p40, p38, p37, and p36) required for proliferating cell nuclear antigen (PCNA)-dependent processive DNA synthesis catalyzed by DNA polymerase δ or ɛ. Here we report the reconstitution of the RFC complex from its five subunits simultaneously overexpressed in baculovirus-infected insect cells. The purified baculovirus-produced RFC appears to contain equimolar levels of each subunit and was shown to be functionally identical to its native counterpart in ( i ) supporting DNA polymerase δ-catalyzed PCNA-dependent DNA chain elongation; ( ii ) catalyzing DNA-dependent ATP hydrolysis that was stimulated by PCNA and human single-stranded DNA binding protein; ( iii ) binding preferentially to DNA primer ends; and ( iv ) catalytically loading PCNA onto singly nicked circular DNA and catalytically removing PCNA from these DNA molecules.

Authors 10

Jinsong Cai (first)
Frank Uhlmann (additional)
Emma Gibbs (additional)
Hernan Flores-Rozas (additional)
Chee-Gun Lee (additional)
Barbara Phillips (additional)
Jeff Finkelstein (additional)
Nina Yao (additional)
Michael O’Donnell (additional)
Jerard Hurwitz (additional)

References 43 Referenced 80

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R Peoples, L Perez-Jurado, Y-K Nung, P Kaplan, U Franke Am J Hum Genet 58, 1370–1373 (1996). / Am J Hum Genet by Peoples R (1996)
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10.1126/science.1598572

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:43 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 3:20 p.m.)
Indexed	4 weeks ago (July 30, 2025, 8:22 p.m.)
Issued	28 years, 9 months ago (Nov. 12, 1996)
Published	28 years, 9 months ago (Nov. 12, 1996)
Published Online	28 years, 9 months ago (Nov. 12, 1996)
Published Print	28 years, 9 months ago (Nov. 12, 1996)

Funders 0

None

BibTeX

@article{Cai_1996, title={Reconstitution of human replication factor C from its five subunits in baculovirus-infected insect cells}, volume={93}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.93.23.12896}, DOI={10.1073/pnas.93.23.12896}, number={23}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Cai, Jinsong and Uhlmann, Frank and Gibbs, Emma and Flores-Rozas, Hernan and Lee, Chee-Gun and Phillips, Barbara and Finkelstein, Jeff and Yao, Nina and O’Donnell, Michael and Hurwitz, Jerard}, year={1996}, month=nov, pages={12896–12901} }

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