Ubiquitin-Binding Domains in Y-Family Polymerases Regulate Translesion Synthesis

Bienko, Magda; Green, Catherine; Crosetto, Nicola; Rudolf, Fabian; Zapart, Grzegorz; Coull, Barry J.; Kannouche, Patricia; Wider, Gerhard; Peter, Matthias; Lehmann, Alan R.; Hofmann, Kay; Đikić, Ivan

doi:10.1126/science.1120615

Cited by 641 publications

(935 citation statements)

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“…Second, the high levels of monoubiquitinated PCNA (due to the loss of USP1) may result in the recruitment and competition of other error-prone TLS polymerases (such as Rev3-Rev7, Rev1, polκ and polι) with polη for undamaged and damaged DNA templates. A recent study has identified two ubiquitin-binding domains (UBM and UBZ) that are evolutionarily conserved in all Y-family TLS polymerases (Rev1, polκ, polι and polη) 12 . These domains are required by polη and polι (and possibly other Y-family TLS polymerases) for interaction with monoubiquitinated PCNA.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, a model has emerged in which polη binds to monoubiquitinated PCNA and ensures accurate (error-free) replicative bypass of UV lesions. However, other (error-prone) TLS polymerases (such as polι and Rev1) have recently been shown to rely on monoubiquitinated PCNA for their function 12,13 . How cells limit PCNA monoubiquitination and the unwanted deployment of polη and/or other error-prone TLS polymerases in the absence or presence of extrinsic DNA damage during the synthesis of DNA in S phase is not known.…”

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confidence: 99%

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Regulation of monoubiquitinated PCNA by DUB autocleavage

et al. 2006

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Monoubiquitination is a reversible post-translational protein modification that has an important regulatory function in many biological processes, including DNA repair. Deubiquitinating enzymes (DUBs) are proteases that are negative regulators of monoubiquitination, but little is known about their regulation and contribution to the control of conjugated-substrate levels. Here, we show that the DUB ubiquitin specific protease 1 (USP1) deubiquitinates the DNA replication processivity factor, PCNA, as a safeguard against error-prone translesion synthesis (TLS) of DNA. Ultraviolet (UV) irradiation inactivates USP1 through an autocleavage event, thus enabling monoubiquitinated PCNA to accumulate and to activate TLS. Significantly, the site of USP1 cleavage is immediately after a conserved internal ubiquitin-like diglycine (Gly-Gly) motif. This mechanism is reminiscent of the processing of precursors of ubiquitin and ubiquitin-like modifiers by DUBs. Our results define a regulatory mechanism for protein ubiquitination that involves the signal-induced degradation of an inhibitory DUB.Monoubiquitination is a highly regulated process that is conserved in all eukaryotes 1,2 and controls a broad range of cellular functions, including DNA repair. Protein monoubiquitination is a reversible post-translational event that can be influenced by the opposing activities of a ubiquitin E3 ligase and a deubiquitinating enzyme (DUB), similar to the regulation of protein phosphorylation by kinases and phosphatases 3,4 . Protein monoubiquitination regulates the rescue of stalled DNA replication forks -an important cellular process required for cell survival 5 . The E2 ubiquitin conjugating enzyme RAD6 and the E3 ligase RAD18 are conserved in both yeast and human and they coordinate and activate the monoubiquitination of PCNA in response to UV damage or stalled replication forks [6][7][8] . Recent studies have shown that polη, a specialized TLS polymerase, is recruited to the replication fork through a specific interaction with monoubiquitinated PCNA 7 . The deployment of TLS polymerases during replication ensures timely bypass of the diverse DNA lesions encountered by the replication fork. Although many TLS polymerases are intrinsically mutagenic, polη allows replication past UV-damaged bases with high fidelity 9-11 . Thus, a model has emerged in which polη binds to monoubiquitinated PCNA and ensures accurate (error-free) replicative bypass of UV lesions. However, other (error-prone) TLS polymerases (such as polι and Rev1) have recently been shown to rely on monoubiquitinated PCNA for their function 12,13 . How cells limit PCNA monoubiquitination and the unwanted deployment of polη and/or other error-prone TLS polymerases in the absence or presence of extrinsic DNA damage during the synthesis of DNA in S phase is not known.In humans, protein deubiquitination is controlled by a family of approximately 95 distinct DUB enzymes 14,15 but the function of most of these proteins is unknown. DUBs are cysteine proteases that cleave ubiquitin fro...

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Section: Discussionmentioning

confidence: 99%

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Regulation of monoubiquitinated PCNA by DUB autocleavage

et al. 2006

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“…PCNA is monoubiquinated at lysine residue 164 in human cells after UV-irradiation, and it has been proposed that this modification may mediate the exchange between replicative and TLS polymerases when the replication fork arrests at a lesion 93 . Accordingly, TLS polymerases bind to monoubiquitinated PCNA through two types of motif: a PCNA interacting peptide (PIP box) that binds directly to PCNA 94 and an ubiquitin-binding motif that is required for their bypass function 95 . The role of PCNA in SHM was analysed using mice bearing a PCNA mutation (Lys164Arg) that is known to inhibit its monoubiquitination and therefore its interaction with TLS polymerases.…”

Section: Polη the Study Of Patients Affected By The Xeroderma Pigmenmentioning

confidence: 99%

DNA polymerases in adaptive immunity

Weill

Reynaud

2008

Nat Rev Immunol

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PrefaceTo cope with an unpredictable variety of potential pathogenic insults, the immune system must generate an enormous diversity of recognition structures, and it does so by making stepwise modifications of key genetic loci in each lymphoid cell. This proceeds through the action of lymphoid-specific proteins acting together with the general DNA-repair machinery of the cell. Strikingly, these general mechanisms are usually diverted from their normal functions, being used in rather atypical ways in order to privilege diversity over accuracy. In this Review, we focus on the contribution of a set of DNA polymerases discovered in the last decade to these unique DNA transactions.

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“…Mono-ubiquitination of PCNA increased its affinity for polη, so that their interaction could be detected in cell extracts [25,26]. It was shown subsequently that not only polη, but also polι, Rev1 and polκ have novel ubiquitinbinding domains (UBDs), and that at least in the case of polη and polι (and likely for polκ and Rev1 also), the polymerases were able to bind to ubiquitin [30]. Thus, the combination of binding to ubiquitinated PCNA via both the PIP motif and the UBDs strengthens the interactions between the polymerases and PCNA, facilitating their recruitment to the stalled fork and facilitating the polymerase switch.…”

Section: Recruitment To the Replication Forkmentioning

confidence: 99%

Translesion synthesis in mammalian cells

Lehmann

2006

Experimental Cell Research

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100

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DNA damage blocks the progression of the replication fork. In order to circumvent the damaged bases, cells employ specialised low stringency DNA polymerases, which are able to carry out translesion synthesis (TLS) past different types of damage. The five polymerases used in TLS in human cells have different substrate specificities, enabling them to deal with many different types of damaged bases. PCNA plays a central role in recruiting the TLS polymerases and effecting the polymerase switch from replicative to TLS polymerase. When the fork is blocked PCNA gets ubiquitinated. This increases its affinity for the TLS polymerases, which all have novel ubiquitin-binding motifs, thereby facilitating their engagement at the stalled fork to effect TLS.

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Ubiquitin-Binding Domains in Y-Family Polymerases Regulate Translesion Synthesis

Cited by 641 publications

References 17 publications

Regulation of monoubiquitinated PCNA by DUB autocleavage

Regulation of monoubiquitinated PCNA by DUB autocleavage

DNA polymerases in adaptive immunity

Translesion synthesis in mammalian cells

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