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The structure and function of MCM from archaeal M. Thermoautotrophicum

Nature Structural & Molecular Biology volume 10, pages 160–167 (2003)Cite this article

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Abstract

Eukaryotic chromosomal DNA is licensed for replication precisely once in each cell cycle. The mini-chromosome maintenance (MCM) complex plays a role in this replication licensing. We have determined the structure of a fragment of MCM from Methanobacterium thermoautotrophicum (mtMCM), a model system for eukaryotic MCM. The structure reveals a novel dodecameric architecture with a remarkably long central channel. The channel surface has an unusually high positive charge and binds DNA. We also show that the structure of the N-terminal fragment is conserved for all MCMs proteins despite highly divergent sequences, suggesting a common architecture for a similar task: gripping/remodeling DNA and regulating MCM activity. An mtMCM mutant protein equivalent to a yeast MCM5 (CDC46) protein with the bob1 mutation at its N terminus has only subtle structural changes, suggesting a Cdc7-bypass mechanism by Bob1 in yeast. Yeast bypass experiments using MCM5 mutant proteins support the hypothesis for the bypass mechanism.

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Figure 1: Overall structure of double-hexamer of N-mtMCM.
Figure 2: The fold and assembly of N-mtMCM.
Figure 3: Unique features of the channel of N-mtMCM and comparison of the channels between N-mtMCM, T7gp4 and PCNA.
Figure 4: Central channel of N-mtMCM double-hexamer and DNA binding.
Figure 5: The structural conservation of the N-terminal half of all MCMs and the structure of N-mtMCM mutant protein with a mutation at the conserved position corresponding to the yeast mcm5-bob1 mutation.
Figure 6: Yeast bypass results of mutations of MCM5 at Pro83 supporting the domain-push hypothesis for bob1 bypass mechanism.

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  • 10 February 2003

    Corrected HTML, added footnote

Notes

  1. *Note: In the version of this article initially published online, this paper contained two mistakes. The first mistake is in the legend of Fig. 3b; the correct legend should read: "b, Side view of the N-mtMCM dodecamer showing the predominantly negatively charged (red) outer surface". The second mistake is in the first paragraph of page 5 (third line from the top); the correct sentence should read: "A surface charge calculation shows that the inner surface of the entire channel is strongly positive (Fig. 3c); in contrast, the outside surface is mostly negative (Fig. 3b)". This mistake has been corrected in the HTML and print versions of the article.

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Acknowledgements

We thank S. Harrison for comments on the manuscript, L. Pessoa-Brandão, R. Zhao, D. Li, T. Gould and L. Wilson for assistance and other members of the Chen group for comments and input; R. Zhang at 19id in Argonne National Laboratory (APS) and the staff at 14bmc in APS and X25 and X4A in Brookhaven National Laboratory for assistance in data collection; and the UCHSC X-ray center in Biomolecular Structure Program for support. This work is supported by start-up and cancer-center funds from UCHSC to X.C. and an NIH grant to R.S.

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  1. Department of Biochemistry and Molecular Genetics University of Colorado Health Science Center, University of Colorado Health Science Center, School of Medicine, Denver, 80262, Colorado, USA

    Ryan J. Fletcher, Brooke E. Bishop, Ronald P. Leon, Robert A. Sclafani & Xiaojiang S. Chen

  2. X4A, HHMI beamline, Brookhaven National Laboratory, Upton, 11973-5000, New York, USA

    Craig M. Ogata

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Fletcher, R., Bishop, B., Leon, R. et al. The structure and function of MCM from archaeal M. Thermoautotrophicum. Nat Struct Mol Biol 10, 160–167 (2003). https://doi.org/10.1038/nsb893

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