Selective Condensation Drives Partitioning and Sequential Secretion of Cyst Wall Proteins in Differentiating Giardia lamblia

Controlled secretion of a protective extracellular matrix is required for transmission of the infective stage of a large number of protozoan and metazoan parasites. Differentiating trophozoites of the highly minimized protozoan parasite Giardia lamblia secrete the proteinaceous portion of the cyst wall material (CWM) consisting of three paralogous cyst wall proteins (CWP1–3) via organelles termed encystation-specific vesicles (ESVs). Phylogenetic and molecular data indicate that Diplomonads have lost a classical Golgi during reductive evolution. However, neogenesis of ESVs in encysting Giardia trophozoites transiently provides basic Golgi functions by accumulating presorted CWM exported from the ER for maturation. Based on this “minimal Golgi” hypothesis we predicted maturation of ESVs to a trans Golgi-like stage, which would manifest as a sorting event before regulated secretion of the CWM. Here we show that proteolytic processing of pro-CWP2 in maturing ESVs coincides with partitioning of CWM into two fractions, which are sorted and secreted sequentially with different kinetics. This novel sorting function leads to rapid assembly of a structurally defined outer cyst wall, followed by slow secretion of the remaining components. Using live cell microscopy we find direct evidence for condensed core formation in maturing ESVs. Core formation suggests that a mechanism controlled by phase transitions of the CWM from fluid to condensed and back likely drives CWM partitioning and makes sorting and sequential secretion possible. Blocking of CWP2 processing by a protease inhibitor leads to mis-sorting of a CWP2 reporter. Nevertheless, partitioning and sequential secretion of two portions of the CWM are unaffected in these cells. Although these cysts have a normal appearance they are not water resistant and therefore not infective. Our findings suggest that sequential assembly is a basic architectural principle of protective wall formation and requires minimal Golgi sorting functions

The C-terminal tail of tetraspanin proteins regulates their intracellular distribution in the parasite Trichomonas vaginalis

The parasite Trichomonas vaginalis is the causative agent of trichomoniasis, a prevalent sexually transmitted infection. Here, we report the cellular analysis of T.vaginalis tetraspanin family (TvTSPs). This family of membrane proteins has been implicated in cell adhesion, migration and proliferation in vertebrates. We found that the expression of several members of the family is up-regulated upon contact with vaginal ectocervical cells. We demonstrate that most TvTSPs are localized on the surface and intracellular vesicles and that the C-terminal intracellular tails of surface TvTSPs are necessary for proper localization. Analyses of full-length TvTSP8 and a mutant that lacks the C-terminal tail indicates that surface-localized TvTSP8 is involved in parasite aggregation, suggesting a role for this protein in parasite:parasite interaction.Fil: Cóceres, Verónica Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas ; ArgentinaFil: Alonso, Andrés Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas ; ArgentinaFil: Nievas, Yésica Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas ; ArgentinaFil: Midlej, V.. Universidade Federal do Rio de Janeiro; BrasilFil: Frontera, L.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas ; ArgentinaFil: Benchimol, M.. Universidade Federal do Rio de Janeiro; BrasilFil: Johnson, P. J.. University of California at Los Angeles; Estados UnidosFil: de Miguel, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas ; Argentin

Cultivos intercalares e controle de plantas daninhas em plantios de maracujá-amarelo Intercropping and weed control in yellow passion fruit orchard

Objetivou-se identificar cultivos intercalares e métodos integrados de controle de plantas daninhas em maracujá-amarelo (Passiflora edulis Sims f. flavicarpa Deg.) como fatores que viabilizem a sua produção e rentabilidade. O experimento foi instalado em maio de 1999, conduzido em espaldeira vertical com um fio de arame a 2,0m do solo, no espaçamento de 2,5m x 5,0m, em blocos casualizados, com seis tratamentos e quatro repetições, sendo T1: milho (Zea mays L. - BR 106) como cultura intercalar; T2: feijão (Phaseolus vulgaris L. - cultivar Pérola) como cultura intercalar; T3: feijão-de-porco nas entrelinhas e capina com enxada nas linhas; T4: feijão-de-porco nas entrelinhas e controle químico nas linhas (glifosate a 1,5 kg/ha); T5: planta daninha controlada quimicamente (em toda a parcela com alachlor a 2,8 kg/ha + diuron a 1,2 kg/ha em pré-emergência e glifosate a 1,5 kg/ha em pós-emergência); e T6: testemunha (capina com enxada em área total). Os dados analisados, no período de produção (maio de 1999 a abril de 2000), mostraram que não houve diferenças estatísticas entre os tratamentos para produtividade (indústria), peso médio, comprimento e diâmetro dos frutos, sólidos solúveis totais e acidez. Contudo, houve significância para produtividade total e in natura, com destaque para a utilização do feijão como cultura intercalar, com produtividade do maracujazeiro de 12,82 t/ha. Tanto o milho como o feijão podem ser recomendados como culturas intercalares no primeiro ano de cultivo do maracujá-amarelo. Os herbicidas aplicados em pré e pós-emergência foram economicamente viáveis e não mostraram efeito tóxico sobre as plantas de maracujá-amarelo.<br>This work aimed identifying crops for intercropping and integrated methods for weed control in yellow passion fruit (Passiflora edulis Sims f. flavicarpa Deg.) orchard, that will favor its production and profitability. The experiment was set up in May 1999, with the plants being conducted by a vertical support structure with one wire extended 2.0 m above the soil surface, and distributed in a spacing of 2.5 m x 5.0 m. The experimental design was in randomized blocks, with four replications and six treatments as follows: T1 - maize (Zea mays L. , cv. BR 106) as intercrop; T2 - bean (Phaseolus vulgaris L., cv. Pérola) as intercrop; T3 - pork bean (Canavalia ensiformis L.) in the interspace and hoeing in the plant lines; T4 - pork bean in the interspace and chemical weed control in the plant lines (glyphosate at 1.5 kg./ha); T5 - general chemical weed control (alachlor at 2.8 kg/ha + diuron at 1.2 kg/ha in pre-emergence and glyphosate at 1.5 kg/ha in post-emergence); T6 - control (general hoeing). Data obtained during the harvest period (May 1999 to April 2000) showed no statistical differences among treatments for yield (industry), fruit weight, length and diameter, total soluble solids and total titrable acidity. However, there was a statistical significance for total yield and yield for the fresh fruit market, with best results for bean as intercrop, giving a passion fruit yield of 12.82 t/ha. Both bean and maize should be recommended as intercrops during the first year of passion fruit cycle. The herbicides used both in pre- and post-emergence were economically viable and did not show toxic effects on the yellow passion fruit plants