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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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.).
The authors declare no competing financial interests.
Sequence alignments of GINS subunits. (PDF 445 kb)
Representative electron density map. (PDF 1887 kb)
Superimpositions of A and B domains. (PDF 504 kb)
Coimmunoprecipitation of GINS with other DNA replication proteins. (PDF 308 kb)
Structural interpretations of other yeast GINS mutants. (PDF 878 kb)
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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