CBP-HSF2 structural and functional interplay in Rubinstein-Taybi neurodevelopmental disorder

Rubinstein-Taybi syndrome (RSTS) is a neurodevelopmental disorder with unclear underlying mechanisms. Here, the authors unravel the contribution of a stress-responsive pathway to RSTS where impaired HSF2 acetylation, due to RSTS-associated CBP/EP300 mutations, alters the expression of neurodevelopmental players, in keeping with hallmarks of cell-cell adhesion defects.Patients carrying autosomal dominant mutations in the histone/lysine acetyl transferases CBP or EP300 develop a neurodevelopmental disorder: Rubinstein-Taybi syndrome (RSTS). The biological pathways underlying these neurodevelopmental defects remain elusive. Here, we unravel the contribution of a stress-responsive pathway to RSTS. We characterize the structural and functional interaction between CBP/EP300 and heat-shock factor 2 (HSF2), a tuner of brain cortical development and major player in prenatal stress responses in the neocortex: CBP/EP300 acetylates HSF2, leading to the stabilization of the HSF2 protein. Consequently, RSTS patient-derived primary cells show decreased levels of HSF2 and HSF2-dependent alteration in their repertoire of molecular chaperones and stress response. Moreover, we unravel a CBP/EP300-HSF2-N-cadherin cascade that is also active in neurodevelopmental contexts, and show that its deregulation disturbs neuroepithelial integrity in 2D and 3D organoid models of cerebral development, generated from RSTS patient-derived iPSC cells, providing a molecular reading key for this complex pathology.</p

A nuclear pore sub-complex restricts the propagation of Ty retrotransposons by limiting their transcription

International audienceBeyond their canonical function in nucleocytoplasmic exchanges, nuclear pore complexes (NPCs) regulate the expression of protein-coding genes. Here, we have implemented transcriptomic and molecular methods to specifically address the impact of the NPC on retroelements, which are present in multiple copies in genomes. We report a novel function for the Nup84 complex, a core NPC building block, in specifically restricting the transcription of LTR-retrotransposons in yeast. Nup84 complex-dependent repression impacts both Copia and Gypsy Ty LTR-retrotransposons, all over the S. cerevisiae genome. Mechanistically, the Nup84 complex restricts the transcription of Ty1, the most active yeast retrotransposon, through the tethering of the SUMO-deconjugating enzyme Ulp1 to NPCs. Strikingly, the modest accumulation of Ty1 RNAs caused by Nup84 complex loss-of-function is sufficient to trigger an important increase of Ty1 cDNA levels, resulting in massive Ty1 retrotransposition. Altogether, our study expands our understanding of the complex interactions between retrotransposons and the NPC, and highlights the importance for the cells to keep retrotransposons under tight transcriptional control

Localization of magma injections, hydrothermal alteration, and deformation in a volcanic detachment (Piton des Neiges, La Réunion)

(IF 2,217)International audienceThis contribution aims at understanding how magmatism, hydrothermal alteration, and deformation may have interacted to localize a detachment (a low-angle normal fault) in a basaltic volcano. Piton des Neiges, an inactive volcano of La Réunion Island, has been deeply cut by erosion, allowing its inner structure to be investigated. The deepest unit observed in the edifice is a kilometer-scale plutonic complex, the top of which is intruded by multiple sills. This zone of repeated sill intrusions has been interpreted as a detachment because it displays evidence of hydrothermal alteration in the greenschist facies linked to a brittle-ductile shear deformation. Deformation begins with cataclasis and is followed by mylonitization and chlorite crystallization, then by hydrofracturing and pumpellyite crystallization. Subsequent and post-deformation calcite crystallization occurs in voids such as fractures and vacuoles. Aluminium substitutions in chlorite suggest that the syn-deformation hydrothermal alteration did not exceed 250 °C and peaked in the deformation zone. Comparison of bulk-rock major element analyses of fresh, altered and deformed rocks shows that the zone of sill intrusion and deformation localized increased concentrations of P and K otherwise depleted in the footwall and hangingwall rocks, suggesting that the detachment acted as a trap for fluids.In contradiction with proposed models of volcano spreading, it is apparent that the portion of Piton des Neiges accessible to observation did not deform by creep of a large hydrothermal system or a plutonic complex below its solidus. Instead, the interface between the already cooled plutonic complex and the host rock acted as a brittle failure zone and was repeatedly intruded by magma injections. This localized heat source promoted hydrothermal alteration and low temperature creep in and around the discontinuity. The same process of magmatism-related weakening might occur on active volcanoes; it may, for instance, explain the slow post-eruptive deformation of the Eastern flank of Piton de La Fournaise (the active volcano of La Réunion) observed since the April 2007 eruption

Discovering New Bioactive Neuropeptides in the Striatum Secretome Using in Vivo Microdialysis and Versatile Proteomics S⃞

The striatum, a major component of the brain basal nuclei, is central for planning and executing voluntary movements and undergoes lesions in neurodegenerative disorders such as Huntington disease. To perform highly integrated tasks, the striatum relies on a complex network of communication within and between brain regions with a key role devoted to secreted molecules. To characterize the rat striatum secretome, we combined in vivo microdialysis together with proteomics analysis of trypsin digests and peptidomics studies of native fragments. This versatile approach, carried out using different microdialysis probes and mass spectrometer devices, allowed evidencing with high confidence the expression of 88 proteins and 100 processed peptides. Their secretory pathways were predicted by in silico analysis. Whereas high molecular weight proteins were mainly secreted by the classical mode (94%), low molecular weight proteins equally used classical and non-classical modes (53 and 47%, respectively). In addition, our results suggested alternative secretion mechanisms not predicted by bioinformatics tools. Based on spectrum counting, we performed a relative quantification of secreted proteins and peptides in both basal and neuronal depolarization conditions. This allowed detecting a series of neuropeptide precursors and a 6-fold increase for neurosecretory protein VGF and proenkephalin (PENK) levels. A focused investigation and a long peptide experiment led to the identification of new secreted non-opioid PENK peptides, referred to as PENK 114–133, PENK 239–260, and PENK 143–185. Moreover we showed that injecting synthetic PENK 114–133 and PENK 239–260 into the striatum robustly increased glutamate release in this region. Thus, the combination of microdialysis and versatile proteomics methods shed new light on the secreted protein repertoire and evidenced novel neuropeptide transmitters

CBP-HSF2 structural and functional interplay in Rubinstein-Taybi neurodevelopmental disorder

International audienceAbstract Patients carrying autosomal dominant mutations in the histone/lysine acetyl transferases CBP or EP300 develop a neurodevelopmental disorder: Rubinstein-Taybi syndrome (RSTS). The biological pathways underlying these neurodevelopmental defects remain elusive. Here, we unravel the contribution of a stress-responsive pathway to RSTS. We characterize the structural and functional interaction between CBP/EP300 and heat-shock factor 2 (HSF2), a tuner of brain cortical development and major player in prenatal stress responses in the neocortex: CBP/EP300 acetylates HSF2, leading to the stabilization of the HSF2 protein. Consequently, RSTS patient-derived primary cells show decreased levels of HSF2 and HSF2-dependent alteration in their repertoire of molecular chaperones and stress response. Moreover, we unravel a CBP/EP300-HSF2-N-cadherin cascade that is also active in neurodevelopmental contexts, and show that its deregulation disturbs neuroepithelial integrity in 2D and 3D organoid models of cerebral development, generated from RSTS patient-derived iPSC cells, providing a molecular reading key for this complex pathology