SPRTN protease and SUMOylation coordinate DNA-protein crosslink repair to prevent genome instability

Vaz, Belén; Ruggiano, Annamaria; Popović, Marta; Rodríguez-Berriguete, Gonzalo; Kilgas, Susan; An, Shi; Gs, Higgins; Kiltie, AE; Ramadan, Kristijan

doi:10.1101/2020.02.14.949289

Cited by 4 publications

(3 citation statements)

References 49 publications

(68 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Emerging evidence suggests that SUMO controls the SprT family of proteases in higher eukaryotes. Although the Wss1 ortholog SPRTN lacks an obvious SIM domain, a recent preprint nevertheless reported that SUMO stimulates its proteolytic activity ( Vaz et al, 2020 ). Another Wss1 ortholog, GCNA, has a SUMO-binding domain and is targeted to DPC sites in a SUMO-dependent manner ( Borgermann et al, 2019 ; Dokshin et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

SUMO orchestrates multiple alternative DNA-protein crosslink repair pathways

et al. 2021

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

SUMO orchestrates multiple alternative DNA-protein crosslink repair pathways

et al. 2021

View full text Add to dashboard Cite

“…Recently DNA-activated proteases ( 6 ) and the proteasome have been suggested to be involved in proteolysis by cleaving large DNA–protein cross-links to DNA–peptide cross-links ( 18 , 29 , 30 ). Some DNA-dependent proteases bind to ubiquitin, or small ubiquitin-like modifier (SUMO), indicative of a role for posttranslational modification in proteolysis ( 31 , 32 , 33 ). Ubiquitination is a key step for proteolysis by the nuclear DNA-dependent metalloprotease SPRTN ( 18 ).…”

mentioning

confidence: 99%

Enzymatic bypass of an N6-deoxyadenosine DNA–ethylene dibromide–peptide cross-link by translesion DNA polymerases

Ghodke¹,

Gonzalez-Vasquez²,

Wang

et al. 2021

Journal of Biological Chemistry

View full text Add to dashboard Cite

Unrepaired DNA–protein cross-links, due to their bulky nature, can stall replication forks and result in genome instability. Large DNA–protein cross-links can be cleaved into DNA–peptide cross-links, but the extent to which these smaller fragments disrupt normal replication is not clear. Ethylene dibromide (1,2-dibromoethane) is a known carcinogen that can cross-link the repair protein O 6 -alkylguanine-DNA alkyltransferase (AGT) to the N6 position of deoxyadenosine (dA) in DNA, as well as four other positions in DNA. We investigated the effect of a 15-mer peptide from the active site of AGT, cross-linked to the N6 position of dA, on DNA replication by human translesion synthesis DNA polymerases (Pols) η, ⍳, and κ. The peptide–DNA cross-link was bypassed by the three polymerases at different rates. In steady-state kinetics, the specificity constant ( k cat / K m ) for incorporation of the correct nucleotide opposite to the adduct decreased by 220-fold with Pol κ, tenfold with pol η, and not at all with Pol ⍳. Pol η incorporated all four nucleotides across from the lesion, with the preference dT > dC > dA > dG, while Pol ⍳ and κ only incorporated the correct nucleotide. However, LC-MS/MS analysis of the primer-template extension product revealed error-free bypass of the cross-linked 15-mer peptide by Pol η. We conclude that a bulky 15-mer peptide cross-linked to the N6 position of dA can retard polymerization and cause miscoding but that overall fidelity is not compromised because only correct pairs are extended.

show abstract

“…It is now known that SUMOylation of TOP2 by the ZATT E3 ligase facilitates the direct removal of TOP2-DNA covalent complexes by TDP2, as discussed above (Schellenberg et al, 2017). However, SUMO is also implicated in other relevant pathways which may affect TOP2-DNA covalent complex processing and repair, such as the SPRTN-dependent pathway (Vaz et al, 2020) and the recently described RNF4/ubiquitin-dependent proteasomal pathway. Following the SUMOylation of TOP2 by the E3 SUMO ligase PIAS4, RNF4 is recruited to TOP2 via its SIM domain and polyubiquitinates TOP2 with K48-linked chains, thereby leading to degradation of TOP2 by the proteasome (Sun et al, 2019).…”

Section: Sumoylation In Top2-dna Covalent Complex Repairmentioning

confidence: 98%

Mechanisms to Repair Stalled Topoisomerase II-DNA Covalent Complexes

2021

View full text Add to dashboard Cite

DNA topoisomerases regulate the topological state of DNA, relaxing DNA supercoils and resolving catenanes and knots that result from biological processes such as transcription and replication. DNA topoisomerase II (TOP2) enzymes achieve this by binding DNA and introducing an enzyme-bridged DNA double-strand break (DSB) where each protomer of the dimeric enzyme is covalently attached to the 5' end of the cleaved DNA via an active site tyrosine phosphodiester linkage. The enzyme then passes a second DNA duplex though the DNA break, before religation and release of the enzyme. However, this activity is potentially hazardous to the cell, as failure to complete religation leads to persistent TOP2 protein-DNA covalent complexes which are cytotoxic. Indeed, this property of topoisomerase has been exploited in cancer therapy in the form of topoisomerase poisons which block the religation stage of the reaction cycle, leading to an accumulation of topoisomerase-DNA adducts. A number of parallel cellular processes have been identified that lead to removal of these covalent TOP2-DNA complexes facilitating repair of the resulting protein-free DSB by standard DNA repair pathways. These pathways presumably arose to repair spontaneous stalled or poisoned TOP2-DNA complexes, but understanding their mechanisms also has implications for cancer therapy, particularly resistance to anti-cancer TOP2 poisons and the genotoxic side effects of these drugs. Here we review recent progress in the understanding of the processing to TOP2 DNA covalent complexes., The basic components and mechanisms plus the additional layer of complexity posed by the post-translational modifications that modulate these pathways. SIGNIFICANCE STATEMENTMultiple pathways have been reported for removal and repair of TOP2-DNA covalent complexes to ensure the timely and efficient repair of TOP2-DNA covalent adducts to protect the genome. Post-translational modifications such as ubiquitination and SUMOylation are involved in the regulation of TOP2-DNA complex repair. Small molecule inhibitors of these post translational modifications may help to improve outcomes of TOP2 poison chemotherapy, for example by increasing TOP2 poison cytotoxicity and reducing genotoxicity, but this remains to be determined.

show abstract

SPRTN protease and SUMOylation coordinate DNA-protein crosslink repair to prevent genome instability

Cited by 4 publications

References 49 publications

SUMO orchestrates multiple alternative DNA-protein crosslink repair pathways

SUMO orchestrates multiple alternative DNA-protein crosslink repair pathways

Enzymatic bypass of an N6-deoxyadenosine DNA–ethylene dibromide–peptide cross-link by translesion DNA polymerases

Mechanisms to Repair Stalled Topoisomerase II-DNA Covalent Complexes

Contact Info

Product

Resources

About