Regulation of the 20S proteasome by a novel family of inhibitory proteins

Olshina, Maya A.; Kumar-Deshmukh, Fanindra; Arkind, Galina; Fainer, Irit; Taranavsky, Mark; Hayat, Daniel; Ben‐Dor, Shifra; Ben-Nissqan, G.; Sharon, Michal

doi:10.1101/617415

Cited by 4 publications

(3 citation statements)

References 81 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We think that HDX-MS could be applied to characterize, not only the binding sites of many new proteasome inhibitors, but also to identify their potential allosteric effects on the 20S core complex. The same is true for the many PIPs that regulate proteasome function and are still poorly characterized from a structural point of view 66 . More broadly, this work demonstrates the ability of HDX-MS to investigate dynamic events on megadalton assemblies including ribosomal particles, inflammasomes, and nucleosomes, to name a few.…”

Section: Discussionmentioning

confidence: 92%

Conformational maps of human 20S proteasomes reveal PA28- and immuno-dependent inter-ring crosstalks

et al. 2020

View full text Add to dashboard Cite

Hydrogen-Deuterium eXchange coupled to Mass Spectrometry (HDX-MS) is now common practice in structural biology. However, it is most of the time applied to rather small oligomeric complexes. Here, we report on the use of HDX-MS to investigate conformational differences between the human standard 20S (std20S) and immuno 20S (i20s) proteasomes alone or in complex with PA28αβ or PA28γ activators. Their solvent accessibility is analyzed through a dedicated bioinformatic pipeline including stringent statistical analysis and 3D visualization. These data confirm the existence of allosteric differences between the std20S and i20S at the surface of the α-ring triggered from inside the catalytic β-ring. Additionally, binding of the PA28 regulators to the 20S proteasomes modify solvent accessibility due to conformational changes of the β-rings. This work is not only a proof-of-concept that HDX-MS can be used to get structural insights on large multi-protein complexes in solution, it also demonstrates that the binding of the std20S or i20S subtype to any of its PA28 activator triggers allosteric changes that are specific to this 20S/PA28 pair.

show abstract

Section: Discussionmentioning

confidence: 92%

Conformational maps of human 20S proteasomes reveal PA28- and immuno-dependent inter-ring crosstalks

et al. 2020

View full text Add to dashboard Cite

show abstract

“…The same is true for the many PIPs that regulate proteasome function and are still poorly characterized from a structural point of view 53 . More broadly, this work demonstrates the ability of HDX-MS to investigate dynamic events on megadalton assemblies including ribosomal particles, inflammasomes and nucleosomes, to name a few.…”

Section: And Fig 8b)mentioning

confidence: 92%

Conformational maps of human 20S proteasomes reveal PA28- and immuno-dependent inter-ring crosstalks

Lesné

Locard‐Paulet

Parra

et al. 2020

Preprint

View full text Add to dashboard Cite

Here, we used for the first time Hydrogen-Deuterium eXchange coupled to Mass Spectrometry (HDX-MS) to investigate conformational differences between the human standard 20S (std20S) and immuno 20S (i20s) proteasomes alone or in complex with PA28αβ or PA28γ activators. Their solvent accessibility was analyzed through a dedicated bioinformatic pipeline including stringent statistical analysis and 3D visualization. These data confirmed the existence of allosteric differences between the std20S and i20S at the surface of the α-ring triggered from inside the catalytic β-ring. Additionally, binding of the PA28 regulators to the 20S proteasomes modified solvent accessibility due to conformational changes of the β-rings. This work is not only a proof-of-concept that HDX-MS can be used to get structural insights on large multi-protein complexes in solution, it also demonstrates that the binding of the std20S or i20S subtype to any of its PA28 activator triggers allosteric changes that are specific to this 20S/PA28 pair.

show abstract

“…It is noteworthy that validated candidate hits from this study, namely LAMTOR2, PSMB2 and PSMC1 are principally genes functioning directly or indirectly in the protein homoeostasis network. Interestingly, RBBP9 has also recently been linked to proteasome function 45 . Combined, these data support the idea that relieving proteotoxic stress may be especially important in HNSCC tumours in FA individuals.…”

Section: Viable Lethalmentioning

confidence: 99%

Genome-wide siRNA screens identify RBBP9 function as a potential target in Fanconi anaemia-deficient head-and-neck squamous cell carcinoma

Pai¹,

Roohollahi²,

Rockx³

et al. 2023

Commun Biol

View full text Add to dashboard Cite

Fanconi anaemia (FA) is a rare chromosomal-instability syndrome caused by mutations of any of the 22 known FA DNA-repair genes. FA individuals have an increased risk of head-and-neck squamous-cell-carcinomas (HNSCC), often fatal. Systemic intolerance to standard cisplatin-based protocols due to somatic-cell hypersensitivity underscores the urgent need to develop novel therapies. Here, we performed unbiased siRNA screens to unveil genetic interactions synthetic-lethal with FA-pathway deficiency in FA-patient HNSCC cell lines. We identified based on differential-lethality scores between FA-deficient and FA-proficient cells, next to common-essential genes such as PSMC1, PSMB2, and LAMTOR2, the otherwise non-essential RBBP9 gene. Accordingly, low dose of the FDA-approved RBBP9-targeting drug Emetine kills FA-HNSCC. Importantly both RBBP9-silencing as well as Emetine spared non-tumour FA cells. This study provides a minable genome-wide analyses of vulnerabilities to address treatment challenges in FA-HNSCC. Our investigation divulges a DNA-cross-link-repair independent lead, RBBP9, for targeted treatment of FA-HNSCCs without systemic toxicity.

show abstract

Regulation of the 20S proteasome by a novel family of inhibitory proteins

Abstract: The protein degradation machinery plays a critical role in the maintenance of cellular homeostasis, preventing the accumulation of damaged or misfolded proteins and controlling the

Cited by 4 publications

References 81 publications

Conformational maps of human 20S proteasomes reveal PA28- and immuno-dependent inter-ring crosstalks

Conformational maps of human 20S proteasomes reveal PA28- and immuno-dependent inter-ring crosstalks

Conformational maps of human 20S proteasomes reveal PA28- and immuno-dependent inter-ring crosstalks

Genome-wide siRNA screens identify RBBP9 function as a potential target in Fanconi anaemia-deficient head-and-neck squamous cell carcinoma

Contact Info

Product

Resources

About