A5033713969- RNA modifications and cancer
- RNA Research and Splicing
- Ubiquitin and proteasome pathways
- Advanced Proteomics Techniques and Applications
- RNA and protein synthesis mechanisms
- Heat shock proteins research
- Genomics and Chromatin Dynamics
- Endoplasmic Reticulum Stress and Disease
- Enzyme Structure and Function
- Genetics and Neurodevelopmental Disorders
- RNA regulation and disease
- DNA Repair Mechanisms
- Bioinformatics and Genomic Networks
- Chromosomal and Genetic Variations
- Cancer-related Molecular Pathways
- Mass Spectrometry Techniques and Applications
- Nuclear Structure and Function
- Fungal and yeast genetics research
- Neurogenetic and Muscular Disorders Research
- Biotin and Related Studies
- Nuclear Receptors and Signaling
- Monoclonal and Polyclonal Antibodies Research
- Herpesvirus Infections and Treatments
- Metabolomics and Mass Spectrometry Studies
- Cellular transport and secretion
Centre National de la Recherche Scientifique
2002-2021
Université de Montpellier
2011-2021
Centre de Recherche en Biologie cellulaire de Montpellier
2012-2017
University of Dundee
2006-2011
Wellcome Trust
2008-2011
Institut de Génétique Moléculaire de Montpellier
2002-2011
Cell and Gene Therapy Catapult
2008
The identification of interaction partners in protein complexes is a major goal cell biology. Here we present reliable affinity purification strategy to identify specific interactors that combines quantitative SILAC-based mass spectrometry with characterization common contaminants binding matrices (bead proteomes). This can be applied either tagged fusion or endogenous complexes, illustrated here using the well-characterized SMN complex as model. GFP used tag choice because it shows minimal...
RNA-binding proteins of the L7Ae family are at heart many essential ribonucleoproteins (RNPs), including box C/D and H/ACA small nucleolar RNPs, U4 nuclear RNP, telomerase, messenger RNPs coding for selenoproteins. In this study, we show that Nufip its yeast homologue Rsa1 key components machinery assembles these RNPs. We observed bind several tether them to other core in immature particles. Surprisingly, also link assembling with AAA + adenosine triphosphatases hRvb1 hRvb2 Hsp90 chaperone...
Immuno-precipitation (IP) experiments using MS provide a sensitive and accurate way of characterising protein complexes their response to regulatory mechanisms. Differences in stoichiometry can be determined as well the reliable identification specific binding partners. The quality control IP interaction studies has its basis biology that is being observed. Is unusual genuine novelty, or an experimental irregularity? Antibodies solid matrices used these techniques isolate not only target...
The box C/D small nucleolar RNPs (snoRNPs) are essential for the processing and modification of rRNA. core proteins restructured during human U3 snoRNP biogenesis; however, molecular basis this is unclear. Here we show that U8 also restructured, suggesting may occur with all snoRNPs. We have characterized four novel biogenesis factors (BCD1, NOP17, NUFIP, TAF9) which, along ATPases TIP48 TIP49, likely to be involved in formation pre-snoRNP. analyzed vitro protein-protein interactions between...
The morphology and composition of subnuclear organelles, such as Cajal bodies (CBs), nucleoli, other nuclear bodies, is dynamic can change in response to a variety cell stimuli, including stress. We show that UV-C irradiation disrupts CBs alters the distribution specific subset CB components. effect on differs from previously reported effects transcription inhibitors. demonstrate mechanism underlying mediated, at least part, by PA28γ (proteasome activator subunit γ). presence...
In vitro, assembly of box C/D small nucleolar ribonucleoproteins (snoRNPs) involves the sequential recruitment core proteins to snoRNAs. vivo, however, factors are required (NUFIP, BCD1, and HSP90–R2TP complex), it is unknown whether a similar scheme applies. this paper, we describe systematic quantitative stable isotope labeling by amino acids in cell culture proteomic experiments crystal structure protein Snu13p/15.5K bound fragment factor Rsa1p/NUFIP. This revealed several unexpected...
HSP90 (Heat Shock Protein 90) is an essential chaperone involved in the last folding steps of client proteins. It has many clients, and these are often recognized through specific adaptors. Recently, conserved R2TP complex was identified as a key co-chaperone. Current evidences indicate that HSP90/R2TP system assembles multi-molecular protein complexes. Strikingly, comprise basic machineries gene expression: (1) nuclear RNA polymerases; (2) snoRNPs, to produce ribosomes; (3) mTOR Complex 1...
The HSP90/R2TP quaternary chaperone assembles key cellular machines, including the three nuclear RNA polymerases and many non-coding RNPs. Here, we characterized associated to R2TP found that it binds partners co-translationally. Its co-translational interactome further reveals novel potential clients identifies bound only co-translationally, post-translationally, or both. For pairs of subunits assembling together co-translationally by R2TP, a marginal proportion their mRNAs is co-localized...
The reliable identification of protein interaction partners and how such interactions change in response to physiological or pathological perturbations is a key goal most areas cell biology. Stable isotope labeling with amino acids culture (SILAC)-based mass spectrometry has been shown provide powerful strategy for characterizing complexes identifying specific interactions. Here, we show SILAC can be combined computational methods drawn from the business intelligence field multidimensional...
Cyclin D1, the regulatory subunit for mid-G(1) cyclin-dependent kinases, controls expression of numerous cell cycle genes. A cyclic AMP-responsive element (CRE), located upstream cyclin D1 mRNA start site, integrates mitogenic signals that target CRE-binding factor CREB, which can recruit transcriptional coactivator CREB-binding protein (CBP). We describe an alternative mechanism CREB-driven induction involves ubiquitous POU domain Oct-1. In breast cancer line MCF-7, overexpression Oct-1 or...
Significance The 20S proteasome is a key actor of the control protein levels and integrity in cells. To perform its multiple functions, it works with series regulators, among which nuclear complex called PA28γ. In particular, PA28γ participates regulation cell proliferation dynamics. We describe here characterization protein, PIP30/FAM192A, binds tightly to favors interaction while inhibiting association coilin, central component Cajal bodies. Thus, PIP30/FAM192A critically controls...
The nucleolus is the subnuclear organelle responsible for ribosome subunit biogenesis and can also act as a stress sensor. It forms around clusters of ribosomal DNA (rDNA) mainly organised in three subcompartments, i.e. fibrillar centre, dense component granular component. Here, we describe localisation 21 protein factors to an intranucleolar region different these main called body (INB). These include proteins involved maintenance, turnover, RNA metabolism, chromatin organisation...
Background information . The CBK1 gene of Saccharomyces cerevisiae encodes a protein kinase that is member the NDR (nuclear Dbf2‐related) family kinases, which are involved in morphogenesis and cell proliferation. Previous studies have shown deletion leads to loss polarity formation large aggregates cells. This aggregation phenotype due daughter cell‐specific accumulation transcription factor Ace2p, responsible for genes whose products necessary final separation mother at end division....
ABSTRACT PA28γ (also known as PSME3), a nuclear activator of the 20S proteasome, is involved in degradation several proteins regulating cell growth and proliferation dynamics various bodies, but its precise cellular functions remain unclear. Here, using quantitative FLIM-FRET based microscopy assay monitoring close proximity between nucleosomes living human cells, we show that controls chromatin compaction. We find depletion induces decompaction pericentromeric heterochromatin, which similar...
ABSTRACT PA28γ is a nuclear activator of the 20S proteasome involved in regulation several essential cellular processes, such as cell proliferation, apoptosis, dynamics and stress response. Unlike 19S regulator proteasome, which specifically recognizes ubiquitylated proteins, promotes degradation substrates by an ATP- ubiquitin-independent manner. However its exact mechanisms action are unclear likely to involve additional partners that remain be identified. Here we report identification...
Abstract PA28γ, a nuclear activator of the 20S proteasome, is involved in degradation several proteins regulating cell growth and proliferation dynamics various bodies, but its precise cellular functions remain unclear. Here, using quantitative FLIM-FRET based microscopy assay monitoring close proximity between nucleosomes living human cells, we show that PA28γ controls chromatin compaction. We find depletion induces decompaction pericentromeric heterochromatin, similarly to observed upon...