Crystal Structure of the Human Rad9–Hus1–Rad1 Clamp

Sohn, Sung-Hwa; Cho, Yunje

doi:10.1016/j.jmb.2009.05.028

Cited by 75 publications

(109 citation statements)

References 65 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Whereas PCNA is comprised of three identical subunits, 9-1-1 is a heterotrimeric sliding clamp (18)(19)(20). This fact reflects the different protein partners the two clamps engage and the distinct roles these complexes play in coordinating DNA processing.…”

mentioning

confidence: 99%

“…Though structural snapshots are available for the individual components of such assemblies (PCNA [Protein Data Bank (PDB) ID code: 1VYM], human 9-1-1 [3GGR]) (Fig. S1) and for two binary complexes [FEN1/DNA (3Q8L) and FEN1/PCNA (1UL1)] (3,16,(18)(19)(20)28), the larger ternary assemblies present Author contributions: J.Q.-A., S.E.T., J.A.T., P.K.C., E.N., and I.I. designed research; J.Q.-A., C.Y., X.X., S.E.T., M.-S.T., and I.I.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Repair complexes of FEN1 endonuclease, DNA, and Rad9-Hus1-Rad1 are distinguished from their PCNA counterparts by functionally important stability

Querol-Audí

Cheng

et al. 2012

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

View full text Add to dashboard Cite

Processivity clamps such as proliferating cell nuclear antigen (PCNA) and the checkpoint sliding clamp Rad9/Rad1/Hus1 (9-1-1) act as versatile scaffolds in the coordinated recruitment of proteins involved in DNA replication, cell-cycle control, and DNA repair. Association and handoff of DNA-editing enzymes, such as flap endonuclease 1 (FEN1), with sliding clamps are key processes in biology, which are incompletely understood from a mechanistic point of view. We have used an integrative computational and experimental approach to define the assemblies of FEN1 with double-flap DNA substrates and either proliferating cell nuclear antigen or the checkpoint sliding clamp 9-1-1. Fully atomistic models of these two ternary complexes were developed and refined through extensive molecular dynamics simulations to expose their conformational dynamics. Clustering analysis revealed the most dominant conformations accessible to the complexes. The cluster centroids were subsequently used in conjunction with single-particle electron microscopy data to obtain a 3D EM reconstruction of the human 9-1-1/FEN1/DNA assembly at 18-Å resolution. Comparing the structures of the complexes revealed key differences in the orientation and interactions of FEN1 and double-flap DNA with the two clamps that are consistent with their respective functions in providing inherent flexibility for lagging strand DNA replication or inherent stability for DNA repair.F lap endonuclease 1 (FEN1) belongs to a class of essential nucleases (the FEN1 5′ nuclease superfamily) present in all domains of life (1). FEN1 catalyzes the endonucleolytic cleavage of bifurcated DNA or RNA structures known as 5′ flaps. These 5′ flaps are generated during lagging strand DNA synthesis or during long-patch base excision repair. The FEN1 substrates are in fact double-flap DNA (dfDNA) with DNA on the opposite side of the 5′ flap, forming a single nucleotide 3′ flap when bound to the enzyme (2, 3). By removing the 5′ ssDNA or RNA flap from such substrates, FEN1 produces a single nicked product that could be sealed by the subsequent action of a DNA ligase (4). Consistent with its crucial role in DNA replication and repair, FEN1 is highly expressed in all proliferative tissues, and its activity is key for the maintenance of genomic integrity (5). FEN1 has been identified as a cancer susceptibility gene, and mutations in it have been linked to a number of genetic diseases, such as EM map myotonic dystrophy, Huntington disease, several ataxias, fragile X syndrome, and cancer (6-10).The nuclease activity of FEN1 can be stimulated by association with processivity clamps such as proliferating cell nuclear antigen (PCNA), which encircle DNA at sites of replication and repair (11)(12)(13). PCNA is a recognized master coordinator of cellular responses to DNA damage and interacts with numerous DNA repair and cell-cycle control proteins. In this capacity, PCNA serves not only as a mobile platform for the attachment of these proteins to DNA but, importantly, plays an active role in the recru...

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

Repair complexes of FEN1 endonuclease, DNA, and Rad9-Hus1-Rad1 are distinguished from their PCNA counterparts by functionally important stability

Querol-Audí

Cheng

et al. 2012

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

View full text Add to dashboard Cite

show abstract

“…These data indicate that Mrad1 plays essential roles in the resistance to UV light, HU and gamma rays, as well as in the S/M and G 2 /M checkpoints. As shown by previous reports (Burtelow et al, 2001;RoosMattjus et al, 2002), as well as 9-1-1 complex crystal structure (Doré et al, 2009;Sohn and Cho, 2009;Xu et al, 2009), Rad1 along with Rad9 and Hus1 in a trimeric checkpoint complex were believed to have similar functions. Indeed as we showed above, many phenotypes such as the hypersensitivity to HU, UV light and gamma rays are similar among Mrad1-deletion, Mrad9-deletion and Mhus1-deletion mouse cells (Fig.…”

Section: Discussionmentioning

confidence: 61%

“…Their protein products form a ring-shaped heterotrimer, named the 9-1-1 complex (Doré et al, 2009;Sohn and Cho, 2009;Xu et al, 2009). It is believed that this complex is important for the functions of DNA repair as well as the activation of cell cycle checkpoints (Shiomi et al, 2002;Bermudez et al, 2003;Ellison and Stillman, 2003).…”

Section: Introductionmentioning

confidence: 99%

Targeted deletion of mouse Rad1 leads to deficient cellular DNA damage responses

Zhang

Liu

et al. 2011

Protein Cell

View full text Add to dashboard Cite

The Rad1 gene is evolutionarily conserved from yeast to human. The fission yeast Schizosaccharomyces pombe Rad1 ortholog promotes cell survival against DNA damage and is required for G 2 /M checkpoint activation. In this study, mouse embryonic stem (ES) cells with a targeted deletion of Mrad1, the mouse ortholog of this gene, were created to evaluate its function in mammalian cells. Mrad1 −/− ES cells were highly sensitive to ultraviolet-light (UV light), hydroxyurea (HU) and gamma rays, and were defective in G 2 /M as well as S/M checkpoints. These data indicate that Mrad1 is required for repairing DNA lesions induced by UV-light, HU and gamma rays, and for mediating G 2 /M and S/M checkpoint controls. We further demonstrated that Mrad1 plays an important role in homologous recombination repair (HRR) in ES cells, but a minor HRR role in differentiated mouse cells.

show abstract

“…The structure of the 9-1-1 complex (Doré et al, 2009;Sohn and Cho, 2009;Xu et al, 2009) is similar to the sliding clamp proliferating cell nuclear antigen protein (PCNA) (Gulbis et al, 1996;Krishna et al, 1994). Hus1 interacts with Rad9 and Rad1 through its N terminal domain and C terminal domain, respectively.…”

mentioning

confidence: 99%