Characterisation of the interaction between WRN, the helicase/exonuclease defective in progeroid Werner's syndrome, and an essential replication factor, PCNA

Rodrı́guez-López, Ana M.; Jackson, Dean A.; Nehlin, Jan O.; Iborra, Francisco J.; Warren, Anna V.; Cox, Lynne S.

doi:10.1016/s0047-6374(02)00131-8

Cited by 48 publications

(46 citation statements)

References 36 publications

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“…Figure 7 shows that PCNA colocalizes partially with both WRN and CAF-1 following HU treatment. This is in agreement with previous reports that PCNA associates with CAF-1 in DNA replication (Moggs et al, 2000), after UV exposure (Green and Almouzni, 2003) process of double-strand breaks repair (Nabatiyan et al, 2006), and that PCNA interacts with WRN during replication (Rodriquez-Lopez et al, 2003). However, how WRN and PCNA coordinately interact with CAF-1 remains to be elucidated.…”

Section: Discussionsupporting

confidence: 92%

The Werner syndrome protein is required for recruitment of chromatin assembly factor 1 following DNA damage

et al. 2006

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The Werner syndrome protein (WRN) and chromatin assembly factor 1 (CAF-1) are both involved in the maintenance of genome stability. In response to DNAdamaging signals, both of these proteins relocate to sites where DNA synthesis occurs. However, the interaction between WRN and CAF-1 has not yet been investigated. In this report, we show that WRN interacts physically with the largest subunit of CAF-1, hp150, in vitro and in vivo. Although hp150 does not alter WRN catalytic activities in vitro, and the chromatin assembly activity of CAF-1 is not affected in the absence of WRN in vivo, this interaction may have an important role during the cellular response to DNA replication fork blockage and/or DNA damage signals. In hp150 RNA-mediated interference (RNAi) knockdown cells, WRN partially formed foci following hydroxyurea (HU) treatment. However, in the absence of WRN, hp150 did not relocate to form foci following exposure to HU and ultraviolet light. Thus, our results demonstrate that WRN responds to DNA damage before CAF-1 and suggest that WRN may recruit CAF-1, via interaction with hp150, to DNA damage sites during DNA synthesis.

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Section: Discussionsupporting

confidence: 92%

The Werner syndrome protein is required for recruitment of chromatin assembly factor 1 following DNA damage

et al. 2006

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“…After the reaction was stopped with formamide dye (80% formamide, 20 mM NaOH, We propose that NEIL1 acts as a cowcatcher in the replication complex when it stalls progression of the replicative polymerase at the lesion site, leading to fork regression, as a result of which the damage in the reannealed duplex can be repaired by NEIL1-initiated BER using the replication proteins. WRN helicase, which interacts with NEIL1 (9), and other replication proteins (43)(44)(45) resolve the chicken-foot after completion of repair (47, 48). 20 mM EDTA, 0.05% bromophenol blue, and 0.05% xylene cyanol), the products were separated on a 20% polyacrylamide gel containing 8 M urea in 1× Tris/borate-EDTA buffer, pH 8.4 (11,58).…”

Section: Methodsmentioning

confidence: 99%

Prereplicative repair of oxidized bases in the human genome is mediated by NEIL1 DNA glycosylase together with replication proteins

Hegde

Hegde²,

Bellot³

et al. 2013

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

133

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Base oxidation by endogenous and environmentally induced reactive oxygen species preferentially occurs in replicating single-stranded templates in mammalian genomes, warranting prereplicative repair of the mutagenic base lesions. It is not clear how such lesions (which, unlike bulky adducts, do not block replication) are recognized for repair. Furthermore, strand breaks caused by base excision from ssDNA by DNA glycosylases, including Nei-like (NEIL) 1, would generate double-strand breaks during replication, which are not experimentally observed. NEIL1, whose deficiency causes a mutator phenotype and is activated during the S phase, is present in the DNA replication complex isolated from human cells, with enhanced association with DNA in S-phase cells and colocalization with replication foci containing DNA replication proteins. Furthermore, NEIL1 binds to 5-hydroxyuracil, the oxidative deamination product of C, in replication protein A-coated ssDNA template and inhibits DNA synthesis by DNA polymerase δ. We postulate that, upon encountering an oxidized base during replication, NEIL1 initiates prereplicative repair by acting as a "cowcatcher" and preventing nascent chain growth. Regression of the stalled replication fork, possibly mediated by annealing helicases, then allows lesion repair in the reannealed duplex. This model is supported by our observations that NEIL1, whose deficiency slows nascent chain growth in oxidatively stressed cells, is stimulated by replication proteins in vitro. Furthermore, deficiency of the closely related NEIL2 alone does not affect chain elongation, but combined NEIL1/2 deficiency further inhibits DNA replication. These results support a mechanism of NEIL1-mediated prereplicative repair of oxidized bases in the replicating strand, with NEIL2 providing a backup function.genome damage repair | replication fork stalling | oxidized base repair at DNA replication fork S everal dozen oxidatively modified, and mostly mutagenic, bases are induced in the genomes of aerobic organisms by endogenous and environmentally induced reactive oxygen species (ROS) (1, 2). For example, 5-hydroxyuracil (5-OHU), a predominant lesion generated by oxidative deamination of C, is mutagenic because of its mispairing with A (3). The bases in the single-stranded (ss) replicating DNA template are particularly prone to oxidation (4); the lack of their repair before replication could fix the mutations. The bulky base adducts if formed in the template strand would block replication and trigger DNA damage-response signaling. In contrast, oxidized bases with minor modifications, which are continuously formed in much higher abundance than the bulky adducts, would mostly allow replication. This raises the question of how these bases are marked for repair before replication to avoid mutagenic consequences. Oxidized base repair in mammalian genomes occurs primarily via the base excision repair (BER) pathway which is initiated with lesion base excision mediated by one of five major DNA glycosylases belonging to the Nth or...

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“…This is supported by the accumulation of stalled replication forks in cells lacking WRN protein (Rodriguez-Lopez et al 2002;Sidorova et al 2008), and it is likely that this reflects a need for the nuclease activity of WRN, since many of the defects observed in WS cells, including reduced proliferative capacity, low S-phase fraction and drug sensitivity, can be corrected by ectopic expression of a bacterial Holliday junction resolvase (Rodriguez-Lopez et al 2007). Recruitment of WRN to stalled replication forks may occur through binding to PCNA via a classical PIP (Lebel et al 1999;Rodriguez-Lopez et al 2003).…”

Section: Wrnmentioning

confidence: 99%

The role of DNA exonucleases in protecting genome stability and their impact on ageing

Mason

Cox

2011

AGE

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Exonucleases are key enzymes involved in many aspects of cellular metabolism and maintenance and are essential to genome stability, acting to cleave DNA from free ends. Exonucleases can act as proofreaders during DNA polymerisation in DNA replication, to remove unusual DNA structures that arise from problems with DNA replication fork progression, and they can be directly involved in repairing damaged DNA. Several exonucleases have been recently discovered, with potentially critical roles in genome stability and ageing. Here we discuss how both intrinsic and extrinsic exonuclease activities contribute to the fidelity of DNA polymerases in DNA replication. The action of exonucleases in processing DNA intermediates during normal and aberrant DNA replication is then assessed, as is the importance of exonucleases in repair of double-strand breaks and interstrand crosslinks. Finally we examine how exonucleases are involved in maintenance of mitochondrial genome stability. Throughout the review, we assess how nuclease mutation or loss predisposes to a range of clinical diseases and particularly ageing.

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Characterisation of the interaction between WRN, the helicase/exonuclease defective in progeroid Werner's syndrome, and an essential replication factor, PCNA

Cited by 48 publications

References 36 publications

The Werner syndrome protein is required for recruitment of chromatin assembly factor 1 following DNA damage

The Werner syndrome protein is required for recruitment of chromatin assembly factor 1 following DNA damage

Prereplicative repair of oxidized bases in the human genome is mediated by NEIL1 DNA glycosylase together with replication proteins

The role of DNA exonucleases in protecting genome stability and their impact on ageing

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