Cryo-EM structures of PP2A:B55 with p107 and Eya3 define substrate recruitment DOI Creative Commons
Sathish K.R. Padi,

Rachel J. Godek,

Wolfgang Peti

et al.

Nature Structural & Molecular Biology, Journal Year: 2025, Volume and Issue: unknown

Published: April 17, 2025

Phosphoprotein phosphatases (PPPs) achieve specificity by binding substrates and regulators using PPP-specific short motifs. Protein phosphatase 2A (PP2A) is a highly conserved that regulates cell signaling tumor suppressor. Here, we use cryo-electron microscopy nuclear magnetic resonance (NMR) spectroscopy to investigate the mechanisms of human p107 substrate Eya3 regulator recruitment PP2A:B55 holoenzyme. We show that, while they associate with B55 common set interaction pockets, mechanism differs distinct from observed for PP2A:B56 other PPPs. also identify core motif in proteins, sequence amongst Eya family. Lastly, NMR-based dephosphorylation assays, demonstrate how directs fidelity through selective specific phosphosites. As orchestrates mitosis DNA damage repair, these data provide roadmap pursuing new avenues therapeutically target this complex individually blocking subset different sites.

Language: Английский

ModelArchive: a deposition database for computational macromolecular structural models DOI Creative Commons
Gerardo Tauriello, Andrew Waterhouse, Juergen Haas

et al.

Journal of Molecular Biology, Journal Year: 2025, Volume and Issue: unknown, P. 168996 - 168996

Published: Feb. 1, 2025

Language: Английский

Citations

3
Cryo-EM structures reveal the PP2A-B55α and Eya3 interaction that can be disrupted by a peptide inhibitor DOI Open Access

Shasha Shi,

Xueni Li,

Christopher P Alderman

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 5, 2025

Abstract We have previously shown that Eya3 recruits PP2A-B55α to dephosphorylate pT58 on Myc, increasing Myc stability and enhancing primary tumor growth of triple-negative breast cancer (TNBC). However, the molecular details how remain unclear. Here we determined cryo-EM structures bound with Eya3, an inhibitory peptide B55i, in its unbound state. These studies demonstrate binds B55α through extended NTD Eya3. The other substrates protein/peptide inhibitors including B55i bind a similar area surface but binding differ. further demonstrated inhibits interaction vitro. expressed plasmid increases decreases protein level TNBC cells, suggesting potential or peptides as therapies for TNBC.

Language: Английский

Citations

0
Defining the Protein Phosphatase 2A (PP2A) Subcomplexes That Regulate FoxO Transcription Factor Localization DOI Creative Commons
Adeline M. Luperchio, Daniel J. Salamango

Cells, Journal Year: 2025, Volume and Issue: 14(5), P. 342 - 342

Published: Feb. 27, 2025

The family of forkhead box O (FoxO) transcription factors regulate cellular processes involved in glucose metabolism, stress resistance, DNA damage repair, and tumor suppression. FoxO transactivation activity is tightly regulated by a complex network signaling pathways post-translational modifications. While it has been well established that phosphorylation promotes cytoplasmic retention inactivation, the mechanism underlying dephosphorylation nuclear translocation less clear. Here, we investigate role protein phosphatase 2A (PP2A) regulating this process. We demonstrate PP2A AMP-activated kinase (AMPK) combine to multiple members following inhibition metabolic or induction oxidative stress. Moreover, chemical inhibitor studies indicate accumulation proteins occurs through export as opposed promoting import previously speculated. Functional, genetic, biochemical identify complexes translocation, binding motif required. Mutating FoxO-PP2A interface enhance diminish alters kinetics accordingly. Together, these shed light on molecular mechanisms provide insights into how regulation integrated with stress-related stimuli.

Language: Английский

Citations

0
PP2A adapter protein IER5 induces dephosphorylation and degradation of MDM2, thereby stabilizing p53 DOI
Taka‐aki Nakada,

Mayuko Koga,

Hiroto Takeuchi

et al.

Cellular Signalling, Journal Year: 2025, Volume and Issue: 131, P. 111739 - 111739

Published: March 11, 2025

Language: Английский

Citations

0
True length of diverse capped RNA sequencing (TLDR-seq): 5′–3′-end sequencing of capped RNAs regardless of 3′-end status DOI Creative Commons
Jamie Auxillos, Arnaud Stigliani,

Christian Vaagensø

et al.

Nucleic Acids Research, Journal Year: 2025, Volume and Issue: 53(6)

Published: March 20, 2025

Abstract Analysis of transcript function is greatly aided by knowledge the full-length RNA sequence. New long-read sequencing enabled Oxford Nanopore and PacBio devices have potential to provide information; however, standard methods still lack ability capture true 5′ ends select for polyadenylated (pA+) transcripts only. Here, we present a method that, utilizing cap trapping 3′-end adapter ligation, sequences between their exact 3′ regardless polyadenylation status without need ribosomal depletion, with characterize length RNAs, if any. The shows high reproducibility, can faithfully detect ends, splice junctions, produces gene-expression estimates that are highly correlated those short-read techniques. We also demonstrate sequence nonadenylated (pA−) including long noncoding promoter upstream transcripts, enhancer cases where pA+ pA− RNAs show preferences different but closely located transcription start sites. Our therefore useful characterization diverse capped species analysis relationships initiation, termination, processing.

Language: Английский

Citations

0
Molecular mechanism of PP2A/B55α phosphatase inhibition by IER5 DOI

Ruili Cao,

Daniel T.D. Jones, Pan Li

et al.

Cell chemical biology, Journal Year: 2025, Volume and Issue: unknown

Published: April 1, 2025

Language: Английский

Citations

0
Cryo-EM structures of PP2A:B55 with p107 and Eya3 define substrate recruitment DOI Creative Commons
Sathish K.R. Padi,

Rachel J. Godek,

Wolfgang Peti

et al.

Nature Structural & Molecular Biology, Journal Year: 2025, Volume and Issue: unknown

Published: April 17, 2025

Phosphoprotein phosphatases (PPPs) achieve specificity by binding substrates and regulators using PPP-specific short motifs. Protein phosphatase 2A (PP2A) is a highly conserved that regulates cell signaling tumor suppressor. Here, we use cryo-electron microscopy nuclear magnetic resonance (NMR) spectroscopy to investigate the mechanisms of human p107 substrate Eya3 regulator recruitment PP2A:B55 holoenzyme. We show that, while they associate with B55 common set interaction pockets, mechanism differs distinct from observed for PP2A:B56 other PPPs. also identify core motif in proteins, sequence amongst Eya family. Lastly, NMR-based dephosphorylation assays, demonstrate how directs fidelity through selective specific phosphosites. As orchestrates mitosis DNA damage repair, these data provide roadmap pursuing new avenues therapeutically target this complex individually blocking subset different sites.

Language: Английский

Citations

0