IER3IP1-mutations cause microcephaly by selective inhibition of ER-Golgi transport.

Mutations in the IER3IP1 (Immediate Early Response-3 Interacting Protein 1) gene can give rise to MEDS1 (Microcephaly with Simplified Gyral Pattern, Epilepsy, and Permanent Neonatal Diabetes Syndrome-1), a severe condition leading to early childhood mortality. The small endoplasmic reticulum (ER)-membrane protein IER3IP1 plays a non-essential role in ER-Golgi transport. Here, we employed secretome and cell-surface proteomics to demonstrate that the absence of IER3IP1 results in the mistrafficking of proteins crucial for neuronal development and survival, including FGFR3, UNC5B and SEMA4D.

Conserved exchange of paralog proteins during neuronal differentiation.

Gene duplication enables the emergence of new functions by lowering the evolutionary pressure that is posed on the ancestral genes. Previous studies have highlighted the role of specific paralog genes during cell differentiation, for example, in chromatin remodeling complexes. It remains unexplored whether similar mechanisms extend to other biological functions and whether the regulation of paralog genes is conserved across species.

Spatiotemporal Control of Lipid Conversion, Actin-Based Mechanical Forces, and Curvature Sensors during Clathrin/AP-1-Coated Vesicle Biogenesis.

Clathrin/adaptor protein-1-coated carriers connect the secretory and the endocytic pathways. Carrier biogenesis relies on distinct protein networks changing membrane shape at the trans-Golgi network, each regulating coat assembly, F-actin-based mechanical forces, or the biophysical properties of lipid bilayers. How these different hubs are spatiotemporally coordinated remains largely unknown.

A high-throughput siRNA screen identifies genes that regulate mannose 6-phosphate receptor trafficking.

The delivery of newly synthesized soluble lysosomal hydrolases to the endosomal system is essential for lysosome function and cell homeostasis. This process relies on the proper trafficking of the mannose 6-phosphate receptors (MPRs) between the trans-Golgi network (TGN), endosomes and the plasma membrane. Many transmembrane proteins regulating diverse biological processes ranging from virus production to the development of multicellular organisms also use these pathways.

Liposome-based assays to study membrane-associated protein networks.

Transport carriers regulate the bidirectional flow of membrane between the compartments of the secretory and endocytic pathways. Their biogenesis relies on the recruitment of a number of cytosolic proteins and protein complexes on specific membrane microdomains with defined protein and lipid compositions. The timely assembly of these cellular machines onto membranes involves multiple protein-protein and protein-lipid interactions and is necessary to select membrane proteins and lipids into nascent carriers, to bend the flat membrane of the donor compartment, to change the shape of this nascent carrier into a tubular-vesicular structure, and to operate its scission from the donor compartment.

Bridging membrane and cytoskeleton dynamics in the secretory and endocytic pathways.

Transport carriers regulate membrane flow between compartments of the secretory and endocytic pathways in eukaryotic cells. Carrier biogenesis is assisted by microtubules, actin filaments and their associated motors that link to membrane-associated coats, adaptors and accessory proteins. We summarize here how the biochemical properties of membranes inform their interactions with cytoskeletal regulators.

Exit from the trans-Golgi network: from molecules to mechanisms.

The trans-Golgi network is a major sorting platform of the secretory pathway from which proteins and lipids, both newly synthesized and retrieved from endocytic compartments, are targeted to different destinations. These sorting processes occur during the formation of pleomorphic tubular-vesicular carriers. The past years have provided insights into basic mechanisms coordinating the spatial and temporal organization of machineries necessary for the segregation of membrane components into distinct microdomains, for the bending, elongation, and fission of corresponding membranes, thus revealing a complex interplay of protein-protein and protein-lipid interactions.

Bidirectional transport between the trans-Golgi network and the endosomal system.

The exchange of proteins and lipids between the trans-Golgi network (TGN) and the endosomal system requires multiple cellular machines, whose activities are coordinated in space and time to generate pleomorphic, tubulo-vesicular carriers that deliver their content to their target compartments. These machines and their associated protein networks are recruited and/or activated on specific membrane domains where they select proteins and lipids into carriers, contribute to deform/elongate and partition membrane domains using the mechanical forces generated by actin polymerization or movement along microtubules. The coordinated action of these protein networks contributes to regulate the dynamic state of multiple receptors recycling between the cell surface, endosomes and the TGN, to maintain cell homeostasis as exemplified by the biogenesis of lysosomes and related organelles, and to establish/maintain cell polarity.

Protein complexes containing CYFIP/Sra/PIR121 coordinate Arf1 and Rac1 signalling during clathrin-AP-1-coated carrier biogenesis at the TGN.

Actin dynamics is a tightly regulated process involved in various cellular events including biogenesis of clathrin-coated, AP-1 (adaptor protein 1)-coated transport carriers connecting the trans-Golgi network (TGN) and the endocytic pathway. However, the mechanisms coordinating coat assembly, membrane and actin remodelling during post-TGN transport remain poorly understood. Here we show that the Arf1 (ADP-ribosylation factor 1) GTPase synchronizes the TGN association of clathrin-AP-1 coats and protein complexes comprising CYFIP (cytoplasmic fragile-X mental retardation interacting protein; Sra, PIR121), a clathrin heavy chain binding protein associated with mental retardation.

Src-dependent repression of ARF6 is required to maintain podosome-rich sealing zones in bone-digesting osteoclasts.

Bone digestion occurs when osteoclasts adhere onto bone surfaces and polarize to form acidic, hydrolase-rich resorption lacunae. For this process, they condense their actin-rich podosomes in tight belts to establish sealing zones, which segregate their basal membranes from those facing resorption lacunae. This polarization process remains poorly understood.

Role for Rab3a in oligodendrocyte morphological differentiation.

Rab3a, a small GTPase important for exocytosis, is uniquely up-regulated as oligodendrocytes enter terminal differentiation and initiate myelin biosynthesis. In this study, we analyze the role of this protein in oligodendrocyte morphological differentiation by using Rab3a overexpression and siRNAi-mediated Rab3a silencing. We found that Rab3a silencing delayed mature oligodendrocyte morphological differentiation but did not interfere with lineage progression of OL progenitors; this is consistent with the high levels of Rab3a expressed by mature oligodendrocytes compared with progenitor cells.

Protein networks supporting AP-3 function in targeting lysosomal membrane proteins.

The AP-3 adaptor complex targets selected transmembrane proteins to lysosomes and lysosome-related organelles. We reconstituted its preferred interaction with liposomes containing the ADP ribosylation factor (ARF)-1 guanosine triphosphatase (GTPase), specific cargo tails, and phosphatidylinositol-3 phosphate, and then we performed a proteomic screen to identify new proteins supporting its sorting function. We identified approximately 30 proteins belonging to three networks regulating either AP-3 coat assembly or septin polymerization or Rab7-dependent lysosomal transport.

Myelin biogenesis: sorting out protein trafficking.

Myelin biogenesis is a complex process involving coordinated exocytosis, endocytosis, mRNA transport and cytoskeletal dynamics. Recent studies indicate that soluble neuronal signals may control the surface expression of proteolipid protein, a process that involves reduced endocytosis and/or increased transport carrier recruitment from an intracellular pool.

A role for Sec8 in oligodendrocyte morphological differentiation.

In the central nervous system, oligodendrocytes synthesize vast amounts of myelin, a multilamellar membrane wrapped around axons that dramatically enhances nerve transmission. A complex apparatus appears to coordinate trafficking of proteins and lipids during myelin synthesis, but the molecular interactions involved are not well understood. We demonstrate that oligodendrocytes express several key molecules necessary for the targeting of transport vesicles to areas of rapid membrane growth, including the exocyst components Sec8 and Sec6 and the multidomain scaffolding proteins CASK and Mint1.