DIS3L2 and LSm proteins are involved in the surveillance of Sm ring-deficient snRNAs DOI Creative Commons
Adriana Roithová, Zuzana Feketová, Štěpánka Vaňáčová

et al.

Nucleic Acids Research, Journal Year: 2020, Volume and Issue: 48(11), P. 6184 - 6197

Published: April 23, 2020

Spliceosomal small nuclear ribonucleoprotein particles (snRNPs) undergo a complex maturation pathway containing multiple steps in the nucleus and cytoplasm. snRNP biogenesis is strictly proofread several quality control checkpoints are placed along pathway. Here, we analyzed fate of RNAs (snRNAs) that unable to acquire ring Sm proteins. We showed snRNAs lacking unstable accumulate P-bodies an LSm1-dependent manner. further provide evidence defective without binding site uridylated at 3' end associate with DIS3L2 3'→5' exoribonuclease LSm Finally, inhibition 5'→3' XRN1 increases association ΔSm DIS3L2, which indicates competition compensation between these two degradation enzymes. Together, degraded by alternative pathways involving either or proteins XRN1.

Language: Английский

DIP-MS: ultra-deep interaction proteomics for the deconvolution of protein complexes DOI Creative Commons
Fabian Frommelt, Andrea Fossati, Federico Uliana

et al.

Nature Methods, Journal Year: 2024, Volume and Issue: 21(4), P. 635 - 647

Published: March 26, 2024

Abstract Most proteins are organized in macromolecular assemblies, which represent key functional units regulating and catalyzing most cellular processes. Affinity purification of the protein interest combined with liquid chromatography coupled to tandem mass spectrometry (AP–MS) represents method choice identify interacting proteins. The composition complex isoforms concurrently present AP sample can, however, not be resolved from a single AP–MS experiment but requires computational inference multiple time- resource-intensive reciprocal experiments. Here we introduce deep interactome profiling by (DIP-MS), combines blue-native-PAGE separation, data-independent acquisition deep-learning-based signal processing resolve sharing same bait experiment. We applied DIP-MS probe organization human prefoldin family complexes, resolving distinct holo- subcomplex variants, complex–complex interactions new subunits that were experimentally validated. Our results demonstrate can reveal proteome modularity at unprecedented depth resolution.

Language: Английский

Citations

14
The RPAP3-Cterminal domain identifies R2TP-like quaternary chaperones DOI Creative Commons
Chloé Maurizy, Marc Quinternet, Yoann Abel

et al.

Nature Communications, Journal Year: 2018, Volume and Issue: 9(1)

Published: May 22, 2018

R2TP is an HSP90 co-chaperone that assembles important macro-molecular machineries. It composed of RPAP3-PIH1D1 heterodimer, which binds the two essential AAA+ATPases RUVBL1/RUVBL2. Here, we resolve structure conserved C-terminal domain RPAP3, and show it directly RUVBL1/RUVBL2 hexamers. The human genome encodes other proteins bearing RPAP3-C-terminal-like domains three containing PIH-like domains. Systematic interaction analyses one RPAP3-like protein, SPAG1, PIH1D2 RUVBL1/2 to form R2TP-like complex termed R2SP. This enriched in testis among 68 potential clients identified, some are expressed others ubiquitous. One substrate liprin-α2, organizes large signaling complexes. Remarkably, R2SP required for liprin-α2 expression assembly complexes, indicating functions quaternary protein folding. Effects stronger at 32 °C, suggesting could help compensating lower temperate testis.

Language: Английский

Citations

73
RPAP3 provides a flexible scaffold for coupling HSP90 to the human R2TP co-chaperone complex DOI Creative Commons

Fabrizio Martino,

Mohinder Pal, Hugo Muñoz-Hernández

et al.

Nature Communications, Journal Year: 2018, Volume and Issue: 9(1)

Published: April 10, 2018

The R2TP/Prefoldin-like co-chaperone, in concert with HSP90, facilitates assembly and cellular stability of RNA polymerase II, complexes PI3-kinase-like kinases such as mTOR. However, the mechanism by which this occurs is poorly understood. Here we use cryo-EM biochemical studies on human R2TP core (RUVBL1-RUVBL2-RPAP3-PIH1D1) reveal distinctive role RPAP3, distinguishing metazoan from smaller yeast equivalent. RPAP3 spans both faces a single RUVBL ring, providing an extended scaffold that recruits clients provides flexible tether for HSP90. A 3.6 Å structure reveals direct interaction C-terminal domain ATPase RUVBL2, necessary but absent yeast. mobile TPR domains map to opposite face associating PIH1D1, mediates client protein recruitment. Thus, platform bringing HSP90 into proximity diverse proteins.

Language: Английский

Citations

70
RUVBL1/RUVBL2 ATPase Activity Drives PAQosome Maturation, DNA Replication and Radioresistance in Lung Cancer DOI Creative Commons
Paul Yenerall, Amit Kumar Das, Shan Wang

et al.

Cell chemical biology, Journal Year: 2019, Volume and Issue: 27(1), P. 105 - 121.e14

Published: Dec. 26, 2019

Language: Английский

Citations

58
RUVBL1–RUVBL2 AAA-ATPase: a versatile scaffold for multiple complexes and functions DOI
María I. Daudén, Andrés López‐Perrote, Óscar Llorca

et al.

Current Opinion in Structural Biology, Journal Year: 2020, Volume and Issue: 67, P. 78 - 85

Published: Oct. 28, 2020

Language: Английский

Citations

57
Assembly of the asymmetric human γ-tubulin ring complex by RUVBL1-RUVBL2 AAA ATPase DOI Creative Commons
Fabian Zimmermann, Marina Serna, Artur Ezquerra

et al.

Science Advances, Journal Year: 2020, Volume and Issue: 6(51)

Published: Dec. 18, 2020

RUVBL1-RUVBL2 regulates assembly of microtubule nucleator γ-tubulin ring complex and allows its recombinant production.

Language: Английский

Citations

53
The structure of the R2T complex reveals a different architecture from the related HSP90 cochaperone R2TP DOI
Alberto Palacios‐Abella, Andrés López‐Perrote, Jasminka Boskovic

et al.

Structure, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 1, 2025

Language: Английский

Citations

1
Upstream ORF-Encoded ASDURF Is a Novel Prefoldin-like Subunit of the PAQosome DOI
Philippe Cloutier, Christian Poitras, Denis Faubert

et al.

Journal of Proteome Research, Journal Year: 2019, Volume and Issue: 19(1), P. 18 - 27

Published: Nov. 18, 2019

The PAQosome is an 11-subunit chaperone involved in the biogenesis of several human protein complexes. We show that ASDURF, a recently discovered upstream open reading frame (uORF) 5′ UTR ASNSD1 mRNA, encodes 12th subunit PAQosome. ASDURF displays significant structural homology to β-prefoldins and assembles with five known subunits prefoldin-like module form heterohexameric complex. A model presented. data presented here provide example eukaryotic uORF-encoded polypeptide whose function not limited cis-acting translational regulation downstream coding sequence highlights importance including alternative ORF products proteomic studies.

Language: Английский

Citations

51
Deep Structural Analysis of RPAP3 and PIH1D1, Two Components of the HSP90 Co-chaperone R2TP Complex DOI Creative Commons
Julien Henri, Marie-Eve Chagot, Maxime Bourguet

et al.

Structure, Journal Year: 2018, Volume and Issue: 26(9), P. 1196 - 1209.e8

Published: July 19, 2018

Language: Английский

Citations

46
Structural mechanism for regulation of the AAA-ATPases RUVBL1-RUVBL2 in the R2TP co-chaperone revealed by cryo-EM DOI Creative Commons
Hugo Muñoz-Hernández, Mohinder Pal, Carlos F. Rodríguez

et al.

Science Advances, Journal Year: 2019, Volume and Issue: 5(5)

Published: May 1, 2019

The human R2TP complex (RUVBL1-RUVBL2-RPAP3-PIH1D1) is an HSP90 co-chaperone required for the maturation of several essential multiprotein complexes, including RNA polymerase II, small nucleolar ribonucleoproteins, and PIKK complexes such as mTORC1 ATR-ATRIP. RUVBL1-RUVBL2 AAA-ATPases are also primary components other INO80 Tip60 remodelers. Despite recent efforts, molecular mechanisms regulating in these remain elusive. Here, we report cryo-EM structures show how access to nucleotide-binding site RUVBL2 coupled binding client recruitment component (PIH1D1) its DII domain. This interaction induces conformational rearrangements that lead destabilization N-terminal segment acts a gatekeeper nucleotide exchange. mechanism couples protein-induced motions domains with accessibility RUVBL1-RUVBL2, it likely general shared RUVBL1-RUVBL2-containing complexes.

Language: Английский

Citations

43