Ubiquitin stimulated reversal of topoisomerase 2 DNA-protein crosslinks by TDP2

Matthew J. Schellenberg; C. Denise Appel; Amanda A. Riccio; Logan R. Butler; Juno M. Krahn; Jenna A. Liebermann; Felipe Cortés-Ledesma; R. Scott Williams

doi:10.1093/NAR/GKAA318

Ubiquitin stimulated reversal of topoisomerase 2 DNA-protein crosslinks by TDP2

Matthew J. Schellenberg, C. Denise Appel, Amanda A. Riccio, Logan R. Butler, Juno M. Krahn, Jenna A. Liebermann, Felipe Cortés-Ledesma, R. Scott Williams

Biochemistry and Molecular Biology

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

Tyrosyl-DNA phosphodiesterase 2 (TDP2) reverses Topoisomerase 2 DNA-protein crosslinks (TOP2-DPCs) in a direct-reversal pathway licensed by ZATT^ZNF451 SUMO2 E3 ligase and SUMOylation of TOP2. TDP2 also binds ubiquitin (Ub), but how Ub regulates TDP2 functions is unknown. Here, we show that TDP2 co-purifies with K63 and K27 poly-Ubiquitinated cellular proteins independently of, and separately from SUMOylated TOP2 complexes. Poly-ubiquitin chains of ≥ Ub₃ stimulate TDP2 catalytic activity in nuclear extracts and enhance TDP2 binding of DNA-protein crosslinks in vitro. X-ray crystal structures and small-angle X-ray scattering analysis of TDP2-Ub complexes reveal that the TDP2 UBA domain binds K63-Ub₃ in a 1:1 stoichiometric complex that relieves a UBA-regulated autoinhibitory state of TDP2. Our data indicates that that poly-Ub regulates TDP2-catalyzed TOP2-DPC removal, and TDP2 single nucleotide polymorphisms can disrupt the TDP2-Ubiquitin interface.

Original language	English (US)
Pages (from-to)	6310-6325
Number of pages	16
Journal	Nucleic acids research
Volume	48
Issue number	11
DOIs	https://doi.org/10.1093/NAR/GKAA318
State	Published - 2020

ASJC Scopus subject areas

Genetics

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title = "Ubiquitin stimulated reversal of topoisomerase 2 DNA-protein crosslinks by TDP2",

abstract = "Tyrosyl-DNA phosphodiesterase 2 (TDP2) reverses Topoisomerase 2 DNA-protein crosslinks (TOP2-DPCs) in a direct-reversal pathway licensed by ZATTZNF451 SUMO2 E3 ligase and SUMOylation of TOP2. TDP2 also binds ubiquitin (Ub), but how Ub regulates TDP2 functions is unknown. Here, we show that TDP2 co-purifies with K63 and K27 poly-Ubiquitinated cellular proteins independently of, and separately from SUMOylated TOP2 complexes. Poly-ubiquitin chains of ≥ Ub3 stimulate TDP2 catalytic activity in nuclear extracts and enhance TDP2 binding of DNA-protein crosslinks in vitro. X-ray crystal structures and small-angle X-ray scattering analysis of TDP2-Ub complexes reveal that the TDP2 UBA domain binds K63-Ub3 in a 1:1 stoichiometric complex that relieves a UBA-regulated autoinhibitory state of TDP2. Our data indicates that that poly-Ub regulates TDP2-catalyzed TOP2-DPC removal, and TDP2 single nucleotide polymorphisms can disrupt the TDP2-Ubiquitin interface.",

author = "Schellenberg, {Matthew J.} and Appel, {C. Denise} and Riccio, {Amanda A.} and Butler, {Logan R.} and Krahn, {Juno M.} and Liebermann, {Jenna A.} and Felipe Cort{\'e}s-Ledesma and Williams, {R. Scott}",

year = "2020",

doi = "10.1093/NAR/GKAA318",

language = "English (US)",

volume = "48",

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AU - Schellenberg, Matthew J.

AU - Appel, C. Denise

AU - Riccio, Amanda A.

AU - Butler, Logan R.

AU - Krahn, Juno M.

AU - Liebermann, Jenna A.

AU - Cortés-Ledesma, Felipe

AU - Williams, R. Scott

PY - 2020

Y1 - 2020

N2 - Tyrosyl-DNA phosphodiesterase 2 (TDP2) reverses Topoisomerase 2 DNA-protein crosslinks (TOP2-DPCs) in a direct-reversal pathway licensed by ZATTZNF451 SUMO2 E3 ligase and SUMOylation of TOP2. TDP2 also binds ubiquitin (Ub), but how Ub regulates TDP2 functions is unknown. Here, we show that TDP2 co-purifies with K63 and K27 poly-Ubiquitinated cellular proteins independently of, and separately from SUMOylated TOP2 complexes. Poly-ubiquitin chains of ≥ Ub3 stimulate TDP2 catalytic activity in nuclear extracts and enhance TDP2 binding of DNA-protein crosslinks in vitro. X-ray crystal structures and small-angle X-ray scattering analysis of TDP2-Ub complexes reveal that the TDP2 UBA domain binds K63-Ub3 in a 1:1 stoichiometric complex that relieves a UBA-regulated autoinhibitory state of TDP2. Our data indicates that that poly-Ub regulates TDP2-catalyzed TOP2-DPC removal, and TDP2 single nucleotide polymorphisms can disrupt the TDP2-Ubiquitin interface.

AB - Tyrosyl-DNA phosphodiesterase 2 (TDP2) reverses Topoisomerase 2 DNA-protein crosslinks (TOP2-DPCs) in a direct-reversal pathway licensed by ZATTZNF451 SUMO2 E3 ligase and SUMOylation of TOP2. TDP2 also binds ubiquitin (Ub), but how Ub regulates TDP2 functions is unknown. Here, we show that TDP2 co-purifies with K63 and K27 poly-Ubiquitinated cellular proteins independently of, and separately from SUMOylated TOP2 complexes. Poly-ubiquitin chains of ≥ Ub3 stimulate TDP2 catalytic activity in nuclear extracts and enhance TDP2 binding of DNA-protein crosslinks in vitro. X-ray crystal structures and small-angle X-ray scattering analysis of TDP2-Ub complexes reveal that the TDP2 UBA domain binds K63-Ub3 in a 1:1 stoichiometric complex that relieves a UBA-regulated autoinhibitory state of TDP2. Our data indicates that that poly-Ub regulates TDP2-catalyzed TOP2-DPC removal, and TDP2 single nucleotide polymorphisms can disrupt the TDP2-Ubiquitin interface.

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Ubiquitin stimulated reversal of topoisomerase 2 DNA-protein crosslinks by TDP2

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