DOI: 10.1073/pnas.0803829105. The architecture of the DNA replication origin recognition complex in <i>Saccharomyces cerevisiae</i>

The architecture of the DNA replication origin recognition complex in Saccharomyces cerevisiae

10.1073/pnas.0803829105

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The origin recognition complex (ORC) is conserved in all eukaryotes. The six proteins of the Saccharomyces cerevisiae ORC that form a stable complex bind to origins of DNA replication and recruit prereplicative complex (pre-RC) proteins, one of which is Cdc6. To further understand the function of ORC we recently determined by single-particle reconstruction of electron micrographs a low-resolution, 3D structure of S. cerevisiae ORC and the ORC–Cdc6 complex. In this article, the spatial arrangement of the ORC subunits within the ORC structure is described. In one approach, a maltose binding protein (MBP) was systematically fused to the N or the C termini of the five largest ORC subunits, one subunit at a time, generating 10 MBP-fused ORCs, and the MBP density was localized in the averaged, 2D EM images of the MBP-fused ORC particles. Determining the Orc1–5 structure and comparing it with the native ORC structure localized the Orc6 subunit near Orc2 and Orc3. Finally, subunit–subunit interactions were determined by immunoprecipitation of ORC subunits synthesized in vitro . Based on the derived ORC architecture and existing structures of archaeal Orc1–DNA structures, we propose a model for ORC and suggest how ORC interacts with origin DNA and Cdc6. The studies provide a basis for understanding the overall structure of the pre-RC.

Bibliography

Chen, Z., Speck, C., Wendel, P., Tang, C., Stillman, B., & Li, H. (2008). The architecture of the DNA replication origin recognition complex in Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences, 105(30), 10326â10331.

Authors 6

Zhiqiang Chen (first)
Christian Speck (additional)
Patricia Wendel (additional)
Chunyan Tang (additional)
Bruce Stillman (additional)
Huilin Li (additional)

References 43 Referenced 64

Dates

Type	When
Created	17 years ago (July 22, 2008, 8:44 p.m.)
Deposited	3 years, 4 months ago (April 12, 2022, 4:07 p.m.)
Indexed	1 month, 2 weeks ago (July 2, 2025, 2:33 p.m.)
Issued	17 years ago (July 29, 2008)
Published	17 years ago (July 29, 2008)
Published Online	17 years ago (July 29, 2008)
Published Print	17 years ago (July 29, 2008)

Funders 0

None

BibTeX

@article{Chen_2008, title={The architecture of the DNA replication origin recognition complex in Saccharomyces cerevisiae}, volume={105}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0803829105}, DOI={10.1073/pnas.0803829105}, number={30}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Chen, Zhiqiang and Speck, Christian and Wendel, Patricia and Tang, Chunyan and Stillman, Bruce and Li, Huilin}, year={2008}, month=jul, pages={10326–10331} }

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