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Structure of the human GINS complex and its assembly and functional interface in replication initiation

Nature Structural & Molecular Biology volume 14, pages 388–396 (2007)Cite this article

Abstract

The eukaryotic GINS complex is essential for the establishment of DNA replication forks and replisome progression. We report the crystal structure of the human GINS complex. The heterotetrameric complex adopts a pseudo symmetrical layered structure comprising two heterodimers, creating four subunit-subunit interfaces. The subunit structures of the heterodimers consist of two alternating domains. The C-terminal domains of the Sld5 and Psf1 subunits are connected by linker regions to the core complex, and the C-terminal domain of Sld5 is important for core complex assembly. In contrast, the C-terminal domain of Psf1 does not contribute to the stability of the complex but is crucial for chromatin binding and replication activity. These data suggest that the core complex ensures a stable platform for the C-terminal domain of Psf1 to act as a key interaction interface for other proteins in the replication-initiation process.

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Figure 1: Overall structure and subunit arrangement of the GINS1Δc complex.
Figure 2: Subunit structures.
Figure 3: Details of the horizontal interfaces.
Figure 4: Details of Sld5 B domain binding interface.
Figure 5: Assembly of GINS subunits.
Figure 6: Replication activity and chromatin binding.

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Acknowledgements

We thank K. Demura and N. Igarashi at the BL-5A beamline of the Photon Factory for assistance with data collection, M. Usui for mass spectrometry measurements, I. Hayashi, T. Hirano and W. Yang for critical reading of the manuscript and M. Izumi, M. Kanemaki, K. Kimura, S. Tada, A. Takemoto, H. Takisawa, K. Yanagi and Y. Zhiying for helpful comments and discussions. The antibody to Xenopus Pol ε p60 was a gift from S. Waga (Osaka University). This work was supported by Grants-in-Aid for Science Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (K.K.) and in part by Solution Oriented Research for Science and Technology from the Japan Science and Technology Agency (K.K. and F.H.).

Author information

Authors and Affiliations

  1. Cellular Physiology Laboratory, RIKEN Discovery Research Institute, 2-1 Hirosawa, Wako, 351-0198, Saitama, Japan

    Katsuhiko Kamada & Fumio Hanaoka

  2. Department of Biology, Graduate School of Science, Osaka University, 1-1 Machikaneyama-chyo, Toyonaka, 560-0043, Osaka, Japan

    Yumiko Kubota, Toshiaki Arata & Yosuke Shindo

  3. Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamada-oka, Suita, 565-0871, Osaka, Japan

    Fumio Hanaoka

Authors
  1. Katsuhiko Kamada
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  2. Yumiko Kubota
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  3. Toshiaki Arata
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  4. Yosuke Shindo
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  5. Fumio Hanaoka
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Contributions

K.K. contributed to structural and molecular biology, biochemistry, manuscript preparation and project direction. Y.K. contributed to molecular biology and manuscript preparation. Y.S. and T.A. performed electron microscopy. F.H. organized the project and prepared the manuscript.

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Correspondence to Katsuhiko Kamada.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Sequence alignments of GINS subunits. (PDF 445 kb)

Supplementary Fig. 2

Representative electron density map. (PDF 1887 kb)

Supplementary Fig. 3

Superimpositions of A and B domains. (PDF 504 kb)

Supplementary Fig. 4

Coimmunoprecipitation of GINS with other DNA replication proteins. (PDF 308 kb)

Supplementary Data 1

Structural interpretations of other yeast GINS mutants. (PDF 878 kb)

Supplementary Data 2

Electron micrographs and cut-open surface of human GINS complex. (PDF 934 kb)

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Kamada, K., Kubota, Y., Arata, T. et al. Structure of the human GINS complex and its assembly and functional interface in replication initiation. Nat Struct Mol Biol 14, 388–396 (2007). https://doi.org/10.1038/nsmb1231

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