Differential Oligomerization of the Deubiquitinases USP25 and USP28 Regulates Their Activities

Sauer, Florian; Klemm, Theresa; Kollampally, Ravi B.; Teßmer, Ingrid; Nair, Radhika K.; Popov, Nikita; Kisker, Caroline

doi:10.1016/j.molcel.2019.02.029

Cited by 45 publications

(49 citation statements)

References 62 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, immunoprecipitation of endogenous USP28 co‐immunoprecipitated endogenous ∆Np63, and vice versa (Fig A). In contrast, antibodies against USP25, a ubiquitin‐specific protease that is structurally very similar to USP28 (; Gersch et al , ; Sauer et al , ), did not co‐immunoprecipitate ∆Np63 although USP25 is readily detectable in A‐431 cells. Correspondingly, antibodies against ∆Np63 did not co‐immunoprecipitate endogenous USP25 (Fig A).…”

Section: Resultsmentioning

confidence: 99%

“…Mutation of this cysteine residue strongly reduced the ability of USP28 to deubiquitylate ∆Np63. Since USP28 is known to homo‐dimerize (Gersch et al , ; Sauer et al , ), overexpression of the catalytic dead isoform may dimerize with endogenous USP28, thereby functioning as a dominant negative mutant. Therefore, inhibiting the catalytic activity of USP28 is likely to be a suitable mechanism to target ∆Np63 in SCC tumours.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Maintaining protein stability of ∆Np63 via USP 28 is required by squamous cancer cells

et al. 2020

View full text Add to dashboard Cite

The transcription factor ∆Np63 is a master regulator of epithelial cell identity and essential for the survival of squamous cell carcinoma (SCC) of lung, head and neck, oesophagus, cervix and skin. Here, we report that the deubiquitylase USP28 stabilizes ∆Np63 and maintains elevated ∆NP63 levels in SCC by counteracting its proteasome‐mediated degradation. Impaired USP28 activity, either genetically or pharmacologically, abrogates the transcriptional identity and suppresses growth and survival of human SCC cells. CRISPR/Cas9‐engineered in vivo mouse models establish that endogenous USP28 is strictly required for both induction and maintenance of lung SCC. Our data strongly suggest that targeting ∆Np63 abundance via inhibition of USP28 is a promising strategy for the treatment of SCC tumours.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Maintaining protein stability of ∆Np63 via USP 28 is required by squamous cancer cells

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Multiangle light scattering. Multiangle light scattering combined with size exclusion chromatography (SEC-MALS) was used to determine the oligomeric state of purified His-flXPA-StrepII in solution, as previously described 65,66 . Briefly, 100 μL of His-flXPA-StrepII at concentrations of 65 μM or 80 μM were injected on a Superdex 200 10/300 column (GE Healthcare) pre-equilibrated with XPA MALS buffer (50 mM HEPES, pH 8.0, 200 mM KCl, 5 mM MgCl 2 , 5 mM DTT, and 10% glycerol) and eluted at a constant flow rate of 0.5 ml/min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Single molecule analysis reveals monomeric XPA bends DNA and undergoes episodic linear diffusion during damage search

et al. 2020

Self Cite

View full text Add to dashboard Cite

Nucleotide excision repair (NER) removes a wide range of DNA lesions, including UVinduced photoproducts and bulky base adducts. XPA is an essential protein in eukaryotic NER, although reports about its stoichiometry and role in damage recognition are controversial. Here, by PeakForce Tapping atomic force microscopy, we show that human XPA binds and bends DNA by ∼60°as a monomer. Furthermore, we observe XPA specificity for the helix-distorting base adduct N-(2'-deoxyguanosin-8-yl)-2-acetylaminofluorene over nondamaged dsDNA. Moreover, single molecule fluorescence microscopy reveals that DNAbound XPA exhibits multiple modes of linear diffusion between paused phases. The presence of DNA damage increases the frequency of pausing. Truncated XPA, lacking the intrinsically disordered N-and C-termini, loses specificity for DNA lesions and shows less pausing on damaged DNA. Our data are consistent with a working model in which monomeric XPA bends DNA, displays episodic phases of linear diffusion along DNA, and pauses in response to DNA damage.

show abstract

“…Furthermore, immunoprecipitation of endogenous USP28 co-immunoprecipitated endogenous ΔNp63, and vice versa (Figure 3B). In contrast, antibodies against USP25, a ubiquitin-specific protease that is structurally very similar to USP28 (Gersch et al, 2019; Sauer et al, 2019), did not co-immunoprecipitate ΔNp63 although USP25 is readily detectable in A431 cells. Conversely, antibodies against ΔNp63 did not co-immunoprecipitate endogenous USP25 (Figure 3B).…”

Section: Resultsmentioning

confidence: 85%

The USP28-ΔNp63 axis is a vulnerability of squamous tumours

Prieto-Garcia

Hartmann

Reissland

et al. 2019

Preprint

View full text Add to dashboard Cite

AbstractThe transcription factor ΔNp63 is a master regulator that establishes epithelial cell identity and is essential for the survival of SCC of lung, head and neck, oesophagus, cervix and skin. Here, we report that the deubiquitylase USP28 stabilizes ΔNp63 protein and maintains elevated ΔNP63 levels in SCC by counteracting its proteasome-mediated degradation. Interference with USP28 activity by genetic means abolishes the transcriptional identity of SCC cells and suppresses growth and survival of human SCC cells. CRISPR/Cas9-engineered mouse models establish that both induction and maintenance of lung SCC strictly depend on endogenous USP28. Targeting ΔNp63 protein abundance in SCC via inhibition of USP28 therefore is a feasible strategy for the treatment of SCC tumours.SignificanceSCC depend on ΔNp63, and its protein abundance is tightly controlled by the ubiquitin proteasome system. Here, we demonstrate the dependence of SCC on USP28 for various human SCC in vitro and in vivo using murine lung tumour models. As inhibitors for deubiquitylases become available, targeting USP28 is a promising therapeutic strategy.

show abstract

Differential Oligomerization of the Deubiquitinases USP25 and USP28 Regulates Their Activities

Abstract: Highlights d USP28 forms an active dimer d USP25 adopts an auto-inhibited tetrameric state d Cancer-associated mutations lead to active USP25 d Neither substrate nor ubiquitin chains disrupt the USP25 tetramer

Cited by 45 publications

References 62 publications

Maintaining protein stability of ∆Np63 via USP 28 is required by squamous cancer cells

Maintaining protein stability of ∆Np63 via USP 28 is required by squamous cancer cells

Single molecule analysis reveals monomeric XPA bends DNA and undergoes episodic linear diffusion during damage search

The USP28-ΔNp63 axis is a vulnerability of squamous tumours

Contact Info

Product

Resources

About