The mechanisms of Hedgehog signalling and its roles in development and disease.

International audienceThe cloning of the founding member of the Hedgehog (HH) family of secreted proteins two decades ago inaugurated a field that has diversified to encompass embryonic development, stem cell biology and tissue homeostasis. Interest in HH signalling increased when the pathway was implicated in several cancers and congenital syndromes. The mechanism of HH signalling is complex and remains incompletely understood. Nevertheless, studies have revealed novel biological insights into this system, including the function of HH lipidation in the secretion and transport of this ligand and details of the signal transduction pathway, which involves Patched 1, Smoothened and GLI proteins (Cubitus interruptus in Drosophila melanogaster), as well as, in vertebrates, primary cilia

Variations in Hedgehog signaling: divergence and perpetuation in Sufu regulation of Gli

The Hedgehog (Hh) proteins play a universal role in metazoan development. Nevertheless, fundamental differences exist between Drosophila and vertebrates in the transduction of the Hh signal, notably regarding the role of primary cilia in mammalian cells. In this issue of Genes & Development, Chen and colleagues (pp. 1910–1928) demonstrate that mouse Suppressor of fused (Sufu) regulates the stability of the transcription factors Gli2 and Gli3 by antagonizing the conserved Gli degradation device mediated by Hib/Spop in a cilia-independent manner

Switch of PKA substrates from Cubitus interruptus to Smoothened in the Hedgehog signalosome complex.

International audienceHedgehog (Hh) signalling is crucial for developmental patterning and tissue homeostasis. In Drosophila, Hh signalling is mediated by a bifunctional transcriptional mediator, called Cubitus interruptus (Ci). Protein Kinase A (PKA)-dependent phosphorylation of the serpentine protein Smoothened (Smo) leads to Ci activation, whereas PKA-dependent phosphorylation of Ci leads to the formation of Ci repressor form. The mechanism that switches PKA from an activator to a repressor is not known. Here we show that Hh signalling activation causes PKA to switch its substrates from Ci to Smo within the Hh signalling complex (HSC). In particular, Hh signalling increases the level of Smo, which then outcompetes Ci for association with PKA and causes a switch in PKA substrate recognition. We propose a new model in which the PKA is constitutively present and active within the HSC, and in which the relative levels of Ci and Smo within the HSC determine differential activation and cellular response to Hh signalling

Evaluating Smoothened as a G-protein-coupled receptor for Hedgehog signalling.

International audienceThe Hedgehog signalling pathway controls numerous developmental processes. In response to Hedgehog, Smoothened (Smo), a seven-pass transmembrane protein, orchestrates pathway signalling and controls transcription factor activation. In the absence of Hedgehog, the receptor Patched indirectly inhibits Smo in a catalytic manner. Many questions surrounding Smo activation and signalling remain. Recent findings in Drosophila and vertebrate systems have provided strong evidence that Smo acts as a G-protein-coupled receptor. We discuss the role and regulation of Smo and reassess similarities between Smo and G-protein-coupled receptors. We also examine recently identified members of the invertebrate and vertebrate Smo signalling cascades that are typical components of G-protein-coupled receptor pathways. Greater understanding of the mechanisms of Smo activation and its signalling pathways will allow implementation of novel strategies to target disorders related to disruption of Hh signalling

Endocytosis of Hedgehog through Dispatched Regulates Long-Range Signaling

SummaryThe proteins of the Hedgehog (Hh) family are secreted proteins exerting short- and long-range control over various cell fates in developmental patterning. The Hh gradient in Drosophila wing imaginal discs consists of apical and basolateral secreted pools, but the mechanisms governing the overall establishment of the gradient remain unclear. We investigated the relative contributions of endocytosis and recycling to control the Hh gradient. We show that, upon its initial apical secretion, Hh is re-internalized. We examined the effect of the resistance-nodulation-division transporter Dispatched (Disp) on long-range Hh signaling and unexpectedly found that Disp is specifically required for apical endocytosis of Hh. Re-internalized Hh is then regulated in a Rab5- and Rab4-dependent manner to ensure its long-range activity. We propose that Hh-producing cells integrate endocytosis and recycling as two instrumental mechanisms contributing to regulate the long-range activity of Hh

Limiteurs de courant supraconducteurs

50-60 Hz electrical engineering represents a considerable potential market for superconductors. Fault current limitation will probably be the first among the possible applications, to find an industrial opening. Exploited features are very specific of the superconductors, namely the large contrast existing between an on-state with high current density and an off-state with high electrical resistivity, obtained as soon as the current reaches a specified level. It thus becomes possible to warrant the grid from any current excursion over a specified value, with considerable impact on the protection and dimensions of conductors and devices, and increased possibilities of interconnexion. The current limiter technology is rather complex ; different structures have been proposed and tested ; they are resistive or inductive, based on low-$T_{\rm c}$ or high-$T_{\rm c}$ superconductors. The most advanced candidate to date is a resistive structure, using conductors made of ultra-fine niobium-titanium filaments in a coppemickel matrix, whose essential features are described hereunder.L'électrotechnique industrielle à 50-60 Hz représente pour les supraconducteurs un marché potentiel considérable. Parmi les applications envisageables, la limitation des courants de défaut sera probablement la première à trouver un essor industriel. Les propriétés mises en jeu sont tout à fait spécifiques des supraconducteurs, à savoir le grand contraste existant entre un état passant à haute densité de courant, et un état bloquant à haute résistivité, obtenu dès que le courant dépasse un seuil donné. Il est ainsi possible de garantir le réseau contre toute excursion du courant au-delà d'une valeur spécifiée, avec un impact important sur sa protection, son dimensionnement, son degré d'interconnexion. La technologie du limiteur de courant est assez complexe ; diverses structures ont été proposées et testées, résistives ou inductives, à base de supraconducteurs BT$_{\rm c}$ ou HT$_{\rm c}$. Le candidat le plus avancé à ce jour est une structure de type résistif, utilisant des conducteurs à filaments ultra-fins de niobium-titane et à matrice de cupronickel, dont on donne ici les caractéristiques essentielles

Dally and Notum regulate the switch between low and high level Hedgehog pathway signalling.

International audienceDuring development, secreted morphogens, such as Hedgehog (Hh), control cell fate and proliferation. Precise sensing of morphogen levels and dynamic cellular responses are required for morphogen-directed morphogenesis, yet the molecular mechanisms responsible are poorly understood. Several recent studies have suggested the involvement of a multi-protein Hh reception complex, and have hinted at an understated complexity in Hh sensing at the cell surface. We show here that the expression of the proteoglycan Dally in Hh-receiving cells in Drosophila is necessary for high but not low level pathway activity, independent of its requirement in Hh-producing cells. We demonstrate that Dally is necessary to sequester Hh at the cell surface and to promote Hh internalisation with its receptor. This internalisation depends on both the activity of the hydrolase Notum and the glycosyl-phosphatidyl-inositol (GPI) moiety of Dally, and indicates a departure from the role of the second glypican Dally-like in Hh signalling. Our data suggest that hydrolysis of the Dally-GPI by Notum provides a switch from low to high level signalling by promoting internalisation of the Hh-Patched ligand-receptor complex

Stability and association of Smoothened, Costal2 and Fused with Cubitus interruptus are regulated by Hedgehog.

International audienceThe mechanisms involved in transduction of the Hedgehog (Hh) signal are of considerable interest to developmental and cancer biologists. Stabilization of the integral membrane protein Smoothened (Smo) at the plasma membrane is a crucial step in Hh signalling but the molecular events immediately downstream of Smo remain to be elucidated. We have shown previously that the transcriptional mediator Cubitus interruptus (Ci) is associated in a protein complex with at least two other proteins, the kinesin-like Costal2 (Cos2) and the serine-threonine kinase Fused (Fu). This protein complex governs the access of Ci to the nucleus. Here we show that, consequent on the stabilization of Smo, Cos2 and Fu are destabilized. Moreover, we find that the Cos2-Fu-Ci protein complex is associated with Smo in membrane fractions both in vitro and in vivo. We also show that Cos2 binding on Smo is necessary for the Hh-dependent dissociation of Ci from this complex. We propose that the association of the Cos2 protein complex with Smo at the plasma membrane controls the stability of the complex and allows Ci activation, eliciting its nuclear translocation