A5087242098- Enzyme Structure and Function
- Heat shock proteins research
- Microtubule and mitosis dynamics
- DNA Repair Mechanisms
- RNA modifications and cancer
- Advanced Electron Microscopy Techniques and Applications
- Mitochondrial Function and Pathology
- Ubiquitin and proteasome pathways
- RNA and protein synthesis mechanisms
- Cancer, Hypoxia, and Metabolism
- PARP inhibition in cancer therapy
- Cancer therapeutics and mechanisms
- Genomics and Phylogenetic Studies
- Medical Imaging Techniques and Applications
- CRISPR and Genetic Engineering
- Microbial Natural Products and Biosynthesis
- Photosynthetic Processes and Mechanisms
- Plant-Microbe Interactions and Immunity
- Genetics, Aging, and Longevity in Model Organisms
- ATP Synthase and ATPases Research
- Genetics and Neurodevelopmental Disorders
- Biochemical and Molecular Research
- Epigenetics and DNA Methylation
- Synthesis and Reactivity of Heterocycles
- Genomic variations and chromosomal abnormalities
ETH Zurich
2023-2024
University of Basel
2022
QB3
2022
University of California, Berkeley
2022
Lawrence Berkeley National Laboratory
2022
Spanish National Cancer Research Centre
2018-2021
Centro de Investigaciones Biológicas Margarita Salas
2012-2018
Consejo Superior de Investigaciones Científicas
2012-2018
CIC bioGUNE
2012
Microtubule nucleation is templated by the γ-tubulin ring complex (γ-TuRC), but its structure deviates from geometry of α-/β-tubulin in microtubule, explaining complex's poor nucleating activity. Several proteins may activate γ-TuRC, mechanisms underlying activation are not known. Here, we determined porcine γ-TuRC purified using CDK5RAP2's centrosomin motif 1 (CM1). We identified an unexpected conformation bound to multiple protein modules containing MZT2, GCP2, and CDK5RAP2, resulting a...
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...
The R2TP complex, comprising the Rvb1p-Rvb2p AAA-ATPases, Tah1p, and Pih1p in yeast, is a specialized Hsp90 co-chaperone required for assembly maturation of multi-subunit complexes. These include small nucleolar ribonucleoproteins, RNA polymerase II, complexes containing phosphatidylinositol-3-kinase-like kinases. structure stoichiometry yeast how it couples to are currently unknown. Here, we determine 3D organization using sedimentation velocity analysis cryo-electron microscopy. 359-kDa...
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...
RuvBL1 and RuvBL2, also known as Pontin Reptin, are AAA+ proteins essential in small nucleolar ribonucloprotein biogenesis, chromatin remodelling, nonsense-mediated messenger RNA decay telomerase assembly, among other functions. They homologous to prokaryotic RuvB, forming single- double-hexameric rings; however, a DNA binding domain II (DII) is inserted within the core. Despite their biological significance, questions remain regarding structure. Here, we report cryo-electron microscopy...
The R2TP (RUVBL1-RUVBL2-RPAP3-PIH1D1) complex, in collaboration with heat shock protein 90 (HSP90), functions as a chaperone for the assembly and stability of complexes, including RNA polymerases, small nuclear ribonucleoprotein particles (snRNPs), phosphatidylinositol 3-kinase (PI3K)-like kinases (PIKKs) such TOR SMG1. PIKK stabilization depends on an additional complex TELO2, TTI1, TTI2 (TTT), whose structure function are poorly understood. cryoelectron microscopy (cryo-EM) human R2TP-TTT...
Polyketide synthases (PKSs) are predominantly microbial biosynthetic enzymes. They assemble highly potent bioactive natural products from simple carboxylic acid precursors. The most versatile families of PKSs organized as assembly lines functional modules. Each module performs one round precursor extension and optional modification, followed by directed transfer the intermediate to next module. While enzymatic domains even modules well understood, higher-order modular architecture PKS...
Summary Microtubule nucleation in cells is templated by the γ-tubulin ring complex (γ-TuRC), a 2.3 MDa multiprotein assembly concentrated at microtubule organizing centers (MTOCs). Current γ-TuRC structures exhibit an open conformation that deviates from geometry of α/β-tubulin microtubule, potentially explaining their low vitro microtubule-nucleating activity. Several proteins have been proposed to activate γ-TuRC, but mechanisms underlying activation are not known. Here, we isolated...
The phosphatidylinositol 3-kinase-like kinases (PIKKs) are large serine-threonine protein with a catalytic domain homologous to the 3-kinase (PI3K). All PIKK family members share general organization comprising conserved C-terminus that contains PI3K domain, which is preceded by N-terminal region made of helical HEAT repeats. In humans, includes six members, play essential roles in various processes including DNA repair and damage signalling (ATM, ATR, DNA-PKcs), control cell growth (mTOR),...
The γ-tubulin ring complex (γ-TuRC) is an essential multiprotein assembly, in which γ-tubulin, GCP2-6, actin, MZT1 and MZT2 form asymmetric cone-shaped structure that provides a template for microtubule nucleation. γ-TuRC recruited to organizing centers (MTOCs), such as centrosomes pre-existing mitotic spindle microtubules, via the evolutionarily-conserved attachment factor NEDD1. NEDD1 contains N-terminal WD40 domain binds C-terminal associates with γ-TuRC. However, structural basis of...
Abstract The R2TP (RUVBL1-RUVBL2-RPAP3-PIH1D1) complex, in collaboration with HSP90, functions as a chaperone for the assembly and stability of protein complexes, including RNA polymerases, snRNPs PI3 kinase-like kinases (PIKK) such TOR SMG1. PIKK stabilisation depends on an additional complex TELO2, TTI1 TTI2 (TTT), whose structure function are poorly understood. We have now determined cryo-EM human R2TP-TTT that together biochemical experiments reveals mechanism recruitment to chaperone....
The number of high-resolution structures protein complexes obtained using cryo-electron microscopy (cryo-EM) is increasing rapidly. Cryo-EM maps large macromolecular frequently contain regions resolved at different resolution levels, and modeling atomic de novo can be difficult for domains determined worse than 5 Å in the absence information from other structures. Here we describe details step-by-step decisions strategy followed to model RUVBL2-binding domain (RBD), a 14 kDa C-terminus RNA...
The R2TP (RUVBL1-RUVBL2-RPAP3-PIH1D1) complex, in collaboration with HSP90, functions as a chaperone for the assembly and stability of protein complexes, including RNA polymerases, snRNPs PI3 kinase-like kinases (PIKK) such TOR SMG1. PIKK stabilisation depends on an additional complex TELO2, TTI1 TTI2 (TTT), whose structure function are poorly understood. We have now determined cryo-EM human R2TP-TTT that together biochemical experiments reveals mechanism recruitment to chaperone....