Sorting of lysosomal proteins

AbstractLysosomes are composed of soluble and transmembrane proteins that are targeted to lysosomes in a signal-dependent manner. The majority of soluble acid hydrolases are modified with mannose 6-phosphate (M6P) residues, allowing their recognition by M6P receptors in the Golgi complex and ensuing transport to the endosomal/lysosomal system. Other soluble enzymes and non-enzymatic proteins are transported to lysosomes in an M6P-independent manner mediated by alternative receptors such as the lysosomal integral membrane protein LIMP-2 or sortilin. Sorting of cargo receptors and lysosomal transmembrane proteins requires sorting signals present in their cytosolic domains. These signals include dileucine-based motifs, DXXLL or [DE]XXXL[LI], and tyrosine-based motifs, YXXØ, which interact with components of clathrin coats such as GGAs or adaptor protein complexes. In addition, phosphorylation and lipid modifications regulate signal recognition and trafficking of lysosomal membrane proteins. The complex interaction of both luminal and cytosolic signals with recognition proteins guarantees the specific and directed transport of proteins to lysosomes

The Mechanisms of Vesicle Budding and Fusion

AbstractGenetic and biochemical analyses of the secretory pathway have produced a detailed picture of the molecular mechanisms involved in selective cargo transport between organelles. This transport occurs by means of vesicular intermediates that bud from a donor compartment and fuse with an acceptor compartment. Vesicle budding and cargo selection are mediated by protein coats, while vesicle targeting and fusion depend on a machinery that includes the SNARE proteins. Precise regulation of these two aspects of vesicular transport ensures efficient cargo transfer while preserving organelle identity

Human Vam6p promotes lysosome clustering and fusion in vivo

Regulated fusion of mammalian lysosomes is critical to their ability to acquire both internalized and biosynthetic materials. Here, we report the identification of a novel human protein, hVam6p, that promotes lysosome clustering and fusion in vivo. Although hVam6p exhibits homology to the Saccharomyces cerevisiae vacuolar protein sorting gene product Vam6p/Vps39p, the presence of a citron homology (CNH) domain at the NH2 terminus is unique to the human protein. Overexpression of hVam6p results in massive clustering and fusion of lysosomes and late endosomes into large (2–3 μm) juxtanuclear structures. This effect is reminiscent of that caused by expression of a constitutively activated Rab7. However, hVam6p exerts its effect even in the presence of a dominant-negative Rab7, suggesting that it functions either downstream of, or in parallel to, Rab7. Data from gradient fractionation, two-hybrid, and coimmunoprecipitation analyses suggest that hVam6p is a homooligomer, and that its self-assembly is mediated by a clathrin heavy chain repeat domain in the middle of the protein. Both the CNH and clathrin heavy chain repeat domains are required for induction of lysosome clustering and fusion. This study implicates hVam6p as a mammalian tethering/docking factor characterized with intrinsic ability to promote lysosome fusion in vivo

A revision and morphological analysis of the Uruguayan species of Stevia (Compositae, Eupatorieae)

Se llevó a cabo una revisión y un análisis morfológico de las especies uruguayas de Stevia (Compositae, Eupatorieae). Caracteres de hojas, inflorescencias, pubescencias y pappus son primordiales para la separación de especies. Steviaentreriensis, S. entreriensis var. minor, y Dissothrixhassleriana se consideraron sinónimos de S. hirsuta, y S. ophryodonta y S. oxylaena sinónimos de S. veronicae. Se designaron lectotipos para los nombres Stevia cinerascens, S. megapotamica, S. linariifolia, S. selloi, S. selloi var. yparacayensis, S. oxylaena y S. veronicae. Stevia burkartii fue excluida de la flora uruguaya. Como resultado, se consideran 10 especies uruguayas para el género: S. aristata, S. cinerascens, S. congesta, S. gratioloides, S. hirsuta, S. multiaristata, S. sabulonis, S. satureiifolia, S. selloi, y S. veronicae. Se proporciona una clave, descripciones, fotografías y mapas de distribución para las especies uruguayas.A revision and a morphological analysis of the Uruguayan species of Stevia (Compositae, Eupatorieae) were performed. Leaf, inflorescence, pubescence and pappus traits were identified as key to separate species. Stevia entreriensis, S. entreriensis var. minor, and Dissothrix hassleriana were considered synonyms of S. hirsuta, and S. ophryodonta and S. oxylaena synonyms of S. veronicae. Lectotypes for the names Stevia cinerascens, S. megapotamica, S. linariifolia, S. selloi, S. selloi var. yparacayensis, S. oxylaena and S. veronicae were designated. Stevia burkartii was excluded from the Uruguayan flora. As a result, 10 Uruguayan species are considered: S. aristata, S. cinerascens, S. congesta, S. gratioloides, S. hirsuta, S. multiaristata, S. sabulonis, S. satureiifolia, S. selloi, and S. veronicae. A key to the Uruguayan species, descriptions, photographs and distribution maps are provided.Fil: Rodríguez Cravero, Juan Facundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Gutierrez, Diego Germán. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Katinas, Liliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. División de Plantas Vasculares; ArgentinaFil: Grossi, Mariana Andrea. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. División de Plantas Vasculares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Bonifacino, Mauricio. Universidad de la República. Facultad de Agricultura; UruguayFil: Marchesi, Eduardo. Universidad de la República. Facultad de Agricultura; Urugua

Role of the mammalian retromer in sorting of the cation-independent mannose 6-phosphate receptor

The cation-independent mannose 6-phosphate receptor (CI-MPR) mediates sorting of lysosomal hydrolase precursors from the TGN to endosomes. After releasing the hydrolase precursors into the endosomal lumen, the unoccupied receptor returns to the TGN for further rounds of sorting. Here, we show that the mammalian retromer complex participates in this retrieval pathway. The hVps35 subunit of retromer interacts with the cytosolic domain of the CI-MPR. This interaction probably occurs in an endosomal compartment, where most of the retromer is localized. In particular, retromer is associated with tubular–vesicular profiles that emanate from early endosomes or from intermediates in the maturation from early to late endosomes. Depletion of retromer by RNA interference increases the lysosomal turnover of the CI-MPR, decreases cellular levels of lysosomal hydrolases, and causes swelling of lysosomes. These observations indicate that retromer prevents the delivery of the CI-MPR to lysosomes, probably by sequestration into endosome-derived tubules from where the receptor returns to the TGN