Shiga toxin induces tubular membrane invaginations for its uptake into cells

Clathrin seems to be dispensable for some endocytic processes and, in several instances, no cytosolic coat protein complexes could be detected at sites of membrane invagination. Hence, new principles must in these cases be invoked to account for the mechanical force driving membrane shape changes. Here we show that the Gb3 (glycolipid)-binding B-subunit of bacterial Shiga toxin induces narrow tubular membrane invaginations in human and mouse cells and model membranes. In cells, tubule occurrence increases on energy depletion and inhibition of dynamin or actin functions. Our data thus demonstrate that active cellular processes are needed for tubule scission rather than tubule formation. We conclude that the B-subunit induces lipid reorganization that favours negative membrane curvature, which drives the formation of inward membrane tubules. Our findings support a model in which the lateral growth of B-subunit-Gb3 microdomains is limited by the invagination process, which itself is regulated by membrane tension. The physical principles underlying this basic cargo-induced membrane uptake may also be relevant to other internalization processes, creating a rationale for conceptualizing the perplexing diversity of endocytic routes.     

Desiccation sensitivity and heat tolerance of Prunus ilicifolia seeds dispersed by American black bears ( Ursus americanus )

Carnivore consumption of fruit is a principle means by which many fleshy-fruited plant species achieve long-distance seed dispersal. We examined carnivore dispersal of hollyleaf cherry ( Prunus ilicifolia ) seeds, specifically assessing the survival, desiccation sensitivity, and germination of seeds found in bear scats. Studies were conducted both in the laboratory and in 2 burn areas in Los Padres National Forest, California. Bear scats containing P. ilicifolia seeds were collected in burned and unburned chaparral. We counted seeds in each scat and noted whether endocarps had tooth punctures or rattled audibly when shaken. For comparative germination trials, we also collected fruits and seeds directly from mature shrubs. In the laboratory, following a cold-moist stratification period, seeds were assessed for germinability. In the field, we compared desiccation rates and germinability of seeds from bear scats and freshly collected seeds. We compared rates of moisture loss and germination for seeds subjected to several different conditions, including 25 °C (room temperature), 30 °C and 65 °C (in the lab), and placement on exposed soil in a burn area (in the field), where midday temperatures were approximately 45 °C but likely much higher by late afternoon. Prunus ilicifolia seeds collected from bear scats were largely undamaged; the vast majority of these seeds germinated. In some cases, germination rate was higher for seeds from scats than for seeds from intact fruits. Several results indicate that desiccation is an important cause of reduced germinability. First, seeds that rattled audibly germinated poorly; and the louder the rattle, the lower the germination percentage. Second, seeds (both fresh and those from bear scats) placed in the field under protective screens had greatly reduced levels of germination (an 84% decline) after only 7 days. Third, seeds dried in the lab, even at relatively moderate temperatures, showed a decline in germination with seed moisture loss. The addition of high temperatures accelerated this decline in germination. We discuss the relevance of heat and desiccation sensitivity of seeds dispersed by bears to successful seed germination in burned and late-seral mesic and xeric chaparral. El consumo de fruta por carnívoros es uno de los principales medios por los que muchas especies de plantas de frutos carnosos logran dispersar sus semillas a través largas distancias. Examinamos la dispersión por carnívoros de las semillas del islay ( Prunus ilicifolia ), un arbusto del chaparral, evaluando concretamente la supervivencia, la sensibilidad a la desecación y la germinación de semillas encontradas en las heces de oso. Estos estudios se llevaron a cabo tanto en el laboratorio como en dos áreas quemadas en Los Padres National Forest, California. Se colectaron heces de oso que contenían semillas de P. ilicifolia en el chaparral quemado y en el no quemado. Contamos las semillas en cada bola fecal y notamos si los endocarpios tenían marcas de dientes y si vibraban audiblemente al agitarse. Para llevar a cabo pruebas comparativas de germinación, también colectamos frutos y semillas directamente de los arbustos maduros. En el laboratorio, después de un período de estratificación fría-húmeda, evaluamos la capacidad de germinación de las semillas. En el campo, comparamos las tasas de desecación y de germinación de semillas de las heces de oso con aquellas de semillas colectadas directamente de los arbustos. Se compararon las tasas de pérdida de humedad y germinación de las semillas sujetas a diversas condiciones, entre ellas 25 °C (temperatura ambiente), 30 °C y 65 °C en el laboratorio, y en el campo sobre suelo expuesto en un área quemada, donde la temperatura al mediodía era ~45 °C pero probablemente mucho más alta por la tarde. En general, las semillas de P. ilicifolia colectadas en las heces de oso no estaban dañadas; la gran mayoría de estas semillas germinaron; en algunos casos la tasa de germinación fue más alta que la de frutos intactos. Varios resultados indicaron que la desecación es una causa importante de la reducción en la capacidad de germinación. Primero, las semillas que vibraban audiblemente germinaron poco, y cuanto más fuerte el sonido, menor la germinación. Segundo, las semillas (tanto las frescas como las colectadas de heces de oso) colocadas en el campo bajo una malla protectora exhibieron niveles de germinación muy reducidos (una disminución del 84%) después de sólo siete días. Tercero, la germinación de las semillas desecadas en el laboratorio, incluso a temperaturas relativamente moderadas, disminuyó con la pérdida de humedad de las semillas. Temperaturas más altas aceleraron esta disminución en la germinación. Discutimos la importancia de la sensibilidad al calor y a la desecación en las semillas dispersadas por los osos para la germinación exitosa en el chaparral mésico y árido, tanto en el chaparral recién quemado como en el que está en las etapas finales de sucesión.     

Rare MTNR1B variants impairing melatonin receptor 1B function contribute to type 2 diabetes

Genome-wide association studies have revealed that common noncoding variants in MTNR1B (encoding melatonin receptor 1B, also known as MT(2)) increase type 2 diabetes (T2D) risk(1,2). Although the strongest association signal was highly significant (P < 1 × 10(-20)), its contribution to T2D risk was modest (odds ratio (OR) of ～1.10-1.15)(1-3). We performed large-scale exon resequencing in 7,632 Europeans, including 2,186 individuals with T2D, and identified 40 nonsynonymous variants, including 36 very rare variants (minor allele frequency (MAF) <0.1%), associated with T2D (OR = 3.31, 95% confidence interval (CI) = 1.78-6.18; P = 1.64 × 10(-4)). A four-tiered functional investigation of all 40 mutants revealed that 14 were non-functional and rare (MAF < 1%), and 4 were very rare with complete loss of melatonin binding and signaling capabilities. Among the very rare variants, the partial- or total-loss-of-function variants but not the neutral ones contributed to T2D (OR = 5.67, CI = 2.17-14.82; P = 4.09 × 10(-4)). Genotyping the four complete loss-of-function variants in 11,854 additional individuals revealed their association with T2D risk (8,153 individuals with T2D and 10,100 controls; OR = 3.88, CI = 1.49-10.07; P = 5.37 × 10(-3)). This study establishes a firm functional link between MTNR1B and T2D risk.     

Postzygotic HRAS and KRAS mutations cause nevus sebaceous and Schimmelpenning syndrome

Nevus sebaceous is a common congenital cutaneous malformation. Affected individuals may develop benign and malignant secondary tumors in the nevi during life. Schimmelpenning syndrome is characterized by the association of nevus sebaceous with extracutaneous abnormalities. We report that of 65 sebaceous nevi studied, 62 (95%) had mutations in the HRAS gene and 3 (5%) had mutations in the KRAS gene. The HRAS c.37G>C mutation, which results in a p.Gly13Arg substitution, was present in 91% of lesions. Nonlesional tissues from 18 individuals had a wild-type sequence, confirming genetic mosaicism. The HRAS c.37G>C mutation was also found in 8 of 8 associated secondary tumors. Mosaicism for HRAS c.37G>C and KRAS c.35G>A mutations was found in two individuals with Schimmelpenning syndrome. Functional analysis of HRAS c.37G>C mutant cells showed constitutive activation of the MAPK and PI3K-Akt signaling pathways. Our results indicate that nevus sebaceous and Schimmelpenning syndrome are caused by postzygotic HRAS and KRAS mutations. These mutations may predispose individuals to the development of secondary tumors in nevus sebaceous.     

Terrestrial isopods from the Oued Laou basin,north-eastern Morocco (Crustacea: Oniscidea), with descriptions of two new genera and seven new species

Thirty-four species of terrestrial isopods (Crustacea: Oniscidea) from the Oued Laou basin, in the Rif area of north-eastern Morocco, are recorded. One genus (Paractenoscia) and seven species (Trichoniscus microphthalmus, Paractenoscia cavernicola, Bathytropa rifensis, Soteriscus gibbosus, S. laouensis, Porcellio pseudornatus, and Eluma praticola) are described as new. The genus Soteriscus, unavailable according to article 13.3 of the International Commission on Zoological Nomenclature (ICZN), is here revalidated by choosing S. gaditanus as type species. Six species (Graeconiscus thermophilus, Ctenoscia minima, Platyarthrus parisii, Porcellio humberti, Porcellio flavocinctus and Eluma caelata) have been fully illustrated to facilitate their identifications. Ctenoscia dorsalis Verhoeff is considered to be a junior synonym of C. minima Dollfus. Porcellio ornatus from southern Spain is also figured for comparison with P. pseudornatus sp. nov. The composition and origin of the oniscidean fauna of the Rif region is briefly discussed.

http://zoobank.org/urn:lsid:zoobank.org:pub:DCBF3103-1463-4A32-9BC0-A4CFE8B762AE