Physical and functional interaction between WRNIP1 and RAD18

Yoshimura, Akari; Seki, Masayuki; Kanamori, Makoto; Tateishi, Seiichiro; Tsurimoto, Toshiki; Tada, Shusuke; Enomoto, Takemi

doi:10.1266/ggs.84.171

Cited by 33 publications

(54 citation statements)

References 34 publications

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“…The fact that WRNIP1 is associated with these HR proteins, even under unperturbed conditions, raises the possibility that they may exist in a single complex ready to safeguard the integrity of the forks whenever they arrested. Previous studies have shown that WRNIP1 binds to forked DNA, which resembles stalled forks (Yoshimura et al, 2009). Our CldU-IP experiments reveal the association of WRNIP1 with replication forks upon replication stress, suggesting that WRNIP1 could be actually recruited to perturbed forks in vivo, also confirming recent observations from iPOND approaches (Dungrawala & Cortez, 2015).…”

Section: Discussionsupporting

confidence: 89%

WRNIP 1 protects stalled forks from degradation and promotes fork restart after replication stress

et al. 2016

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Accurate handling of stalled replication forks is crucial for the maintenance of genome stability. RAD51 defends stalled replication forks from nucleolytic attack, which otherwise can threaten genome stability. However, the identity of other factors that can collaborate with RAD51 in this task is poorly elucidated. Here, we establish that human Werner helicase interacting protein 1 (WRNIP1) is localized to stalled replication forks and cooperates with RAD51 to safeguard fork integrity. We show that WRNIP1 is directly involved in preventing uncontrolled MRE11-mediated degradation of stalled replication forks by promoting RAD51 stabilization on ssDNA. We further demonstrate that replication fork protection does not require the ATPase activity of WRNIP1 that is however essential to achieve the recovery of perturbed replication forks. Loss of WRNIP1 or its catalytic activity causes extensive DNA damage and chromosomal aberrations. Intriguingly, downregulation of the anti-recombinase FBH1 can compensate for loss of WRNIP1 activity, since it attenuates replication fork degradation and chromosomal aberrations in WRNIP1-deficient cells. Therefore, these findings unveil a unique role for WRNIP1 as a replication fork-protective factor in maintaining genome stability.

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Section: Discussionsupporting

confidence: 89%

WRNIP 1 protects stalled forks from degradation and promotes fork restart after replication stress

et al. 2016

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“…7 Moreover, additional in vitro studies revealed that WRNIP1 binds in an ATP-dependent manner to forked DNA that mimics stalled forks. 8 Taken together, these findings strongly support the hypothesis of a specific role of WRNIP1 in the management of perturbed replication forks.…”

supporting

confidence: 71%

WRNIP1: A new guardian of genome integrity at stalled replication forks

Leuzzi

Marabitti

Pichierri

et al. 2016

Molecular & Cellular Oncology

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“…Binding of WRN to Template-Primer DNA WRNIP1 and WRN bind to a variety of DNA substrates in vitro 22,23) , however, in vivo substrates for these proteins are not known. Since WRNIP1 and WRN are reported to stimulate the DNA synthesizing activity of Pold in vitro, 15,17) substrates used to assay DNA polymerase activity of Pold, which mimic the structure of a primer annealed on a template DNA, were employed to examine the biochemical relationships between WRNIP1 and WRN.…”

Section: Resultsmentioning

confidence: 99%

“…22) Furthermore, WRNIP1 was recruited to DNA bound by RAD18 via protein-protein interactions and displaces RAD18 from the DNA. 22) In this context, it seems likely that WRNIP1 is recruited to the RAD18-DNA complex and displaces RAD18, followed by WRNIP1-induced recruitment of Pold and WRN to the DNA originally recognized by RAD6-RAD18. Next, WRN may displace WRNIP1 and DNA helicase activity of WRN may enhance DNA polymerase activity of Pold in the elongation step.…”

Section: Discussionmentioning

confidence: 99%

“…19) 46-Oligomer A3 is partially complementary to 20-oligomer A1. 19) Oligonucleotide 120N2R-40ss was previously reported, [20][21][22] and oligonucleotide 20N2 (3Ј) was 5Ј-GCATC CTCAC CATCA ACTCA-3Ј. The template-primer DNA with a 20-nucleotide double-strand region and 60-nucleotide single-strand region was prepared by annealing 120N2R-40ss and 20N2 (3Ј).…”

Section: Protein Preparationmentioning

confidence: 99%

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Werner Interacting Protein 1 Promotes Binding of Werner Protein to Template-Primer DNA

Kanamori

Seki

Yoshimura

et al. 2011

Biological & Pharmaceutical Bulletin

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Werner syndrome (WS) is a rare autosomal recessive disorder that displays many of the clinical symptoms of normal aging at an early age. From the second decade of life onward, WS patients develop pathologies that resemble many of the traits of normal aging, such as osteoporosis, ocular cataracts, graying and loss of hair, diabetes mellitus, arteriosclerosis, and cancer. 1) WS is caused by mutation of a single gene that encodes a 1432-amino acid protein (WRN).2) WRN possesses multiple enzyme activities, including DNA-dependent ATPase, exonuclease, and DNA helicase activities. [3][4][5] Somatic cells derived from WS patients display subtle replication defects, genomic instability, and altered telomere dynamics, suggesting an important role of WRN in DNA metabolic pathways. [6][7][8][9] This is consistent with observations that a variety of proteins that interact with WRN are involved in DNA replication, repair, recombination, transcription, and maintenance of telomere structure.3) We identified a new protein that interacts with WRN and designated it originally Werner Helicase Interacting Protein (WHIP) 10) and renamed it later as WRN interacting protein 1 (WRNIP1). WRNIP1 belongs to the AAAϩ class of the ATPase family and is conserved from Escherichia (E.) coli to human. 10)Mutation in the gene encoding the budding yeast ortholog of WRNIP1, maintenance of genome stability 1 (MGS1) causes hyper recombination and early aging in yeast cells. 10,11) Genetic analyses using MGS1 mutants revealed that Mgs1p is required to prevent genome instability caused by replication arrest and is not involved in the repair of DNA lesions.12) In addition, overproduction of Mgs1p is lethal or very toxic when it is combined with mutations in the genes that encode proteins involved in DNA replication, such as replication protein A, replication factor C, proliferating cell nuclear antigen and DNA polymerase d (Pold). 13)Mgs1p physically and functionally interacts in vivo with budding yeast Pol31, the second subunit of Pold.14) Human WRNIP1 also interacts directly with human Pold and stimulates the DNA synthesizing activity of Pold by increasing initiation frequency of DNA synthesis from primers annealed on template DNA (template-primer DNA).15) In this context, it is of note that WRN interacts with Pold 16) and stimulates the DNA synthesizing activity of Pold by increasing the length of synthesized DNA.17) Since WRNIP1 interacts with WRN, 10,18) WRNIP1 and WRN may interact with templateprimer DNA in a cooperative fashion to regulate the activity of Pold. However, the biochemical relationships between WRNIP1 and WRN at the template-primer DNA remain elusive.In this study, the biochemical relationships between purified WRNIP1 and WRN on template-primer DNA were analyzed. MATERIALS AND METHODS Protein PreparationPreparation of baculovirus for the expression of human WRNIP1 was described previously. 15)High-5 cells infected with recombinant virus encoding FLAG-WRNIP1 were cultured for 3 d and collected by centrifugation. Cells were lysed with...

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Physical and functional interaction between WRNIP1 and RAD18

Cited by 33 publications

References 34 publications

WRNIP 1 protects stalled forks from degradation and promotes fork restart after replication stress

WRNIP 1 protects stalled forks from degradation and promotes fork restart after replication stress

WRNIP1: A new guardian of genome integrity at stalled replication forks

Werner Interacting Protein 1 Promotes Binding of Werner Protein to Template-Primer DNA

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