Crystal structure of the human GINS complex

Choi, Jung Min; Lim, Hyun‐Suk; Kim, Jeong Joo; Song, Ok-Kyu; Cho, Yunje

doi:10.1101/gad.1548107

Cited by 58 publications

(69 citation statements)

References 22 publications

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“…Since archaeal GINS complexes resemble their eukaryotic counterparts in architecture (29), the possibility exists that SsoGINS binds DNA in the same manner. Crystallographic and electron microscopic studies on eukaryotic GINS have revealed various sizes for the central channel, raising the possibility that the protein complex is flexible in structure to permit changes under various conditions (8,42,(45)(46)(47). Therefore, it may be speculated that SsoGINS is capable of undergoing conformational changes upon binding to DNA or when passing DNA into its central channel.…”

Section: Discussionmentioning

confidence: 99%

The Sulfolobus solfataricus GINS Complex Stimulates DNA Binding and Processive DNA Unwinding by Minichromosome Maintenance Helicase

Lang

Huang

2015

J Bacteriol

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GINS is a key component of the eukaryotic Cdc45-minichromosome maintenance (MCM)-GINS (CMG) complex, which unwinds duplex DNA at the moving replication fork. Archaeal GINS complexes have been shown to stimulate the helicase activity of their cognate MCM mainly by elevating its ATPase activity. Here, we report that GINS from the thermoacidophilic crenarchaeon Sulfolobus solfataricus (SsoGINS) is capable of DNA binding and binds preferentially to single-stranded DNA (ssDNA) over double-stranded DNA (dsDNA). Notably, SsoGINS binds more strongly to dsDNA with a 5= ssDNA tail than to dsDNA with a 3= tail and more strongly to an ssDNA fragment blocked at the 3= end than to one at the 5= end with a biotinstreptavidin (SA) complex, suggesting the ability of the protein complex to slide in a 5=-to-3= direction along ssDNA. DNAbound SsoGINS enhances DNA binding by SsoMCM. Furthermore, SsoGINS increases the helicase activity of SsoMCM. However, the ATPase activity of SsoMCM is not affected by SsoGINS. Our results suggest that SsoGINS facilitates processive DNA unwinding by SsoMCM by enhancing the binding of the helicase to DNA. We propose that SsoGINS stabilizes the interaction of SsoMCM with the replication fork and moves along with the helicase as the fork progresses. IMPORTANCEGINS is a key component of the eukaryotic Cdc45-MCM-GINS complex, a molecular motor that drives the unwinding of DNA in front of the replication fork. Archaea also encode GINS, which interacts with MCM, the helicase. But how archaeal GINS serves its role remains to be understood. In this study, we show that GINS from the hyperthermophilic archaeon Sulfolobus solfataricus is able to bind to DNA and slide along ssDNA in a 5=-to-3= direction. Furthermore, Sulfolobus GINS enhances DNA binding by MCM, which slides along ssDNA in a 3=-to-5= direction. Taken together, these results suggest that Sulfolobus GINS may stabilize the interaction of MCM with the moving replication fork, facilitating processive DNA unwinding. Chromosomal DNA replication is a highly regulated process involving a large number of essential and optional protein factors (1, 2). Efficient DNA unwinding is an essential prerequisite for the duplication of the genetic material. In Eukarya, the minichromosome maintenance (MCM) complex comprising six subunits (MCM2 to MCM7) is the replicative helicase. However, the heterohexameric MCM itself shows little helicase activity (3, 4), and two accessory factors, i.e., the tetrameric GINS complex (Sld5, Pfs1, Psf2, and Psf3) and Cdc45, are required to activate MCM (5). The complex of MCM, Cdc45, and GINS, referred to as the CMG complex, is considered to be the active form of eukaryotic helicase (5, 6). The eukaryotic GINS complex is able to bind to the origin of replication and move along with the replication fork (6-11). Cdc45 is also required for the progression of the replication fork (12, 13). Three-dimensional reconstructions of the overall arrangement of the Drosophila CMG based on single-particle electron microscopy reveal that Cdc...

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Section: Discussionmentioning

confidence: 99%

The Sulfolobus solfataricus GINS Complex Stimulates DNA Binding and Processive DNA Unwinding by Minichromosome Maintenance Helicase

Lang

Huang

2015

J Bacteriol

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“…Actually, rotary-shadowed electron microscopic images of the Xenopus GINS complex revealed ring-like structures (7). Furthermore, recent structural studies showed that the crystal structures of the human GINS complex resemble a trapezoid (36) or elliptical shape (37,38), in which the four subunits interact with each other.…”

Section: Discussionmentioning

confidence: 99%

The GINS Complex from Pyrococcus furiosus Stimulates the MCM Helicase Activity

Yoshimochi

Fujikane

Kawanami

et al. 2008

Journal of Biological Chemistry

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Pyrococcus furiosus, a hyperthermophilic Archaea, has homologs of the eukaryotic MCM (mini-chromosome maintenance) helicase and GINS complex. The MCM and GINS proteins are both essential factors to initiate DNA replication in eukaryotic cells. Many biochemical characterizations of the replication-related proteins have been reported, but it has not been proved that the homologs of each protein are also essential for replication in archaeal cells. Here, we demonstrated that the P. furiosus GINS complex interacts with P. furiosus MCM. A chromatin immunoprecipitation assay revealed that the GINS complex is detected preferentially at the oriC region on Pyrococcus chromosomal DNA during the exponential growth phase but not in the stationary phase. Furthermore, the GINS complex stimulates both the ATPase and DNA helicase activities of MCM in vitro. These results strongly suggest that the archaeal GINS is involved in both the initiation and elongation processes of DNA replication in P. furiosus, as observed in eukaryotic cells.

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“…As this work was being reviewed, another study reporting the GINS complex structure with truncated Psf1 by Choi et al was published (19).…”

Section: Methodsmentioning

confidence: 99%

Crystal structure of the GINS complex and functional insights into its role in DNA replication

Chang

Wang

Bermudez

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

120

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The GINS complex, which contains the four subunits Sld5, Psf1, Psf2, and Psf3, is essential for both the initiation and progression of DNA replication in eukaryotes. GINS associates with the MCM2-7 complex and Cdc45 to activate the eukaryotic minichromosome maintenance helicase. It also appears to interact with and stimulate the polymerase activities of DNA polymerase and the DNA polymerase ␣-primase complex. To further understand the functional role of GINS, we determined the crystal structure of the full-length human GINS heterotetramer. Each of the four subunits has a major domain composed of an ␣-helical bundle-like structure. With the exception of Psf1, each of the other subunits has a small domain containing a three-stranded ␤-sheet core. Each full-length protein in the crystal has unstructured regions that are all located on the surface of GINS and are probably involved in its interaction with other replication factors. The four subunits contact each other mainly through ␣-helices to form a ring-like tetramer with a central pore. This pore is partially plugged by a 16-residue peptide from the Psf3 N terminus, which is unique to some eukaryotic Psf3 proteins and is not required for tetramer formation. Removal of these N-terminal 16 residues of Psf3 from the GINS tetramer increases the opening of the pore by 80%, suggesting a mechanism by which accessibility to the pore may be regulated. The structural data presented here indicate that the GINS tetramer is a highly stable complex with multiple flexible surface regions.Cdc45 ͉ DNA helicase ͉ minichromosome maintenance complex ͉ DNA polymerase E ukaryotic DNA replication is controlled by a series of ordered and regulated steps (1-3) that commence with the binding of the six-subunit origin recognition complex (ORC) to replication origins. During the G1 phase of the cell cycle, Cdc6, and Cdt1 are recruited to the origin, and together with ORC, support the loading of the heterohexameric MCM2-7 complex (minichromosome maintenance, MCM) to form the prereplication complex (pre-RC). Although a substantial amount of data suggest that MCM acts as the replicative helicase, MCM present in the pre-RC (as well as isolated MCM) is devoid of helicase activity (summarized in ref. 4). At the G 1 /S transition of the cell cycle, it appears that the MCM helicase activity is activated by a complex and an as yet poorly understood series of modifications that require the action of two protein kinases, DDK (Cdc7-Dbf4) and CDK (cyclin-dependent), as well as the participation of at least eight additional factors, including Mcm10, Cdc45, Dpb11, GINS, synthetic lethal with dpb11 mutant-2 (Sld2), and Sld3 (4). Two of these components, Cdc45 and GINS, appear critical for helicase activation because DNA unwinding is observed (3, 5), concomitant with their loading at origins. In accord with these findings, a complex containing nearstoichiometric levels of MCM, Cdc45, and GINS was isolated from Drosophila and shown to possess DNA helicase activity (6). Studies with Xenopus extracts revealed...

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Crystal structure of the human GINS complex

Cited by 58 publications

References 22 publications

The Sulfolobus solfataricus GINS Complex Stimulates DNA Binding and Processive DNA Unwinding by Minichromosome Maintenance Helicase

The Sulfolobus solfataricus GINS Complex Stimulates DNA Binding and Processive DNA Unwinding by Minichromosome Maintenance Helicase

The GINS Complex from Pyrococcus furiosus Stimulates the MCM Helicase Activity

Crystal structure of the GINS complex and functional insights into its role in DNA replication

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