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Cover image for Vol. 15 Issue 9

Early View (Online Version of Record published before inclusion in an issue)

Edited By: Michael S. Marks, Mark C. P. Marsh, Trina A. Schroer, Tom H. Stevens

Online ISSN: 1600-0854

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  1. 1 - 9
  1. Original Articles

    1. Host PI(3,5)P2 Activity Is Required for Plasmodium berghei Growth During Liver Stage Infection

      Carolina Thieleke-Matos, Mafalda Lopes da Silva, Laura Cabrita-Santos, Cristiana F. Pires, José S. Ramalho, Ognian Ikonomov, Elsa Seixas, Assia Shisheva, Miguel C. Seabra and Duarte C. Barral

      Article first published online: 20 AUG 2014 | DOI: 10.1111/tra.12190

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      The asymptomatic liver stage of malaria infection provides an opportunity for therapeutic intervention to block disease progression and transmission. However, the interaction of Plasmodium parasites with host liver cells remains largely obscure. We found that host PI(3,5)P2 phosphoinositide and its effector TRPML1 are necessary for Plasmodium growth. Hence, we propose that host endocytic vesicles fuse with the parasite vacuole, allowing the exchange of material between the host and the parasite, necessary for parasite replication in the liver.

    2. Eng2 Is a Component of a Dynamic Protein Complex Required for Endocytic Uptake in Fission Yeast

      Javier Encinar del Dedo, Fatima-Zahra Idrissi, Yolanda Arnáiz-Pita, Michael James, Encarnación Dueñas-Santero, Sara Orellana-Muñoz, Francisco del Rey, Vladimir Sirotkin, M. Isabel Geli and Carlos R. Vázquez de Aldana

      Article first published online: 18 AUG 2014 | DOI: 10.1111/tra.12198

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      Eng2 is a cytoplasmic glucanase required for spore release, although it is also present during vegetative growth. Our results indicate that Eng2 is a component of a novel endocytic functional module, which probably couples the endocytic coat to the actin module.

    3. PIKfyve Inhibition Interferes with Phagosome and Endosome Maturation in Macrophages

      Grace H.E. Kim, Roya M. Dayam, Akriti Prashar, Mauricio Terebiznik and Roberto J. Botelho

      Article first published online: 16 AUG 2014 | DOI: 10.1111/tra.12199

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      PIKfyve synthesizes phosphatidylinositol-3,5-bisphosphate, which controls endolysosome function. We examined the requirement of PIKfyve in the macrophage endolysosomal system and its impact on phagosome maturation through gene silencing and pharmacology. Among other observations, PIKfyve activity is important for the timely depletion of phosphatidylinositol-3-phosphate from phagosomes and for efficient phagosome–lysosome fusion. Importantly, PIKfyve inhibition reduced the degradative capacity of phagosomes but did not impact phagosome acidification. We suggest that PIKfyve has direct and indirect roles in phagocytosis and phagosome maturation in macrophages.

    4. AP-1A Controls Secretory Granule Biogenesis and Trafficking of Membrane Secretory Granule Proteins

      Mathilde Bonnemaison, Nils Bäck, Yimo Lin, Juan S. Bonifacino, Richard Mains and Betty Eipper

      Article first published online: 15 AUG 2014 | DOI: 10.1111/tra.12194

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      Peptides and their processing enzymes leave the trans-Golgi network (TGN) in immature secretory granules (SGs). Corticotrope tumor cells were used to explore the role of AP-1A in the generation of secretagogue-responsive mature SGs. Reducing levels of the μ1A subunit of AP-1A resulted in a decrease in TGN cisternae and immature SGs and the appearance of vacuolar structures containing cleavage products unique to the regulated secretory pathway (non-condensing SGs). A twofold reduction in μ1A levels substantially impaired regulated secretion.

    5. Multiple Domains of Tetanus Toxin Direct Entry into Primary Neurons

      Faith C. Blum, William H. Tepp, Eric A. Johnson and Joseph T. Barbieri

      Article first published online: 11 AUG 2014 | DOI: 10.1111/tra.12197

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      Total internal reflection fluorescence (TIRF) microscopy examined tetanus neurotoxin (TeNT) and receptor-binding domain of TeNT (HCR/T) entry into rat primary cortical neurons. TeNT association was independent of membrane depolarization, unaffected by blocking SV cycling and segregated from SVs. In contrast, HCR/T association was increased with membrane depolarization, inhibited by blocking SV cycling and colocalized with SVs. Thus, TeNT enters neurons independent of SVs and TeNT trafficking is directed by regions outside the HCR domain.

  2. Reviews

    1. Ciliopathies: The Trafficking Connection

      Kayalvizhi Madhivanan and Ruben Claudio Aguilar

      Article first published online: 11 AUG 2014 | DOI: 10.1111/tra.12195

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      The receptor-rich, signaling organelle known as primary cilium (PC) has been implicated in a multitude of diseases collectively denominated as ciliopathies. The presence of a defined barrier at its base makes the PC an isolated compartment that needs vesicle trafficking for maintenance and function. This review discusses the major players involved in trafficking to the PC and highlights evidence correlating the severity of the ciliopathy to the trafficking defect involved. Specifically, mild and severe symptoms arise from specific ciliary receptor mislocalization and generalized ciliogenesis defect, respectively.

  3. Toolbox

    1. Immuno- and Correlative Light Microscopy-Electron Tomography Methods for 3D Protein Localization in Yeast

      Muriel Mari, Willie J.C. Geerts and Fulvio Reggiori

      Article first published online: 11 AUG 2014 | DOI: 10.1111/tra.12192

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      Three-dimensional reconstructions with nanoscale resolution in combination with protein subcellular localization are a key investigation tool for the molecular dissection of cellular processes. Here we present two complementary methods optimized for the study of yeast ultrastructure. The first is an immuno-electron tomography (IET) procedure, which allows determining protein distribution patterns on reconstructed organelles. The second is a correlative light microscopy-electron tomography method where compartments containing a specific protein that has been localized through a fluorescent signal are resolved in 3D.

  4. Original Articles

    1. A Role for Exocyst in Maturation and Bactericidal Function of Staphylococci-Containing Endothelial Cell Phagosomes

      Liane Rauch, Kirsten Hennings and Martin Aepfelbacher

      Article first published online: 11 AUG 2014 | DOI: 10.1111/tra.12189

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      Staphylococci that invade human endothelial cells via integrins can be efficiently eliminated in phagolysosomes (PLys). We found that exocyst complex proteins (e.g. YFP-Sec10) and recycling endosome marker Rab11 colocalize on vesicles that interact with maturing endothelial cell phagosomes. Knockdown of exocyst and Rab11 greatly reduced acidification of phagosomes as visualized by pHRodo, a pH sensitive red-fluorescing dye. This effect significantly diminished elimination of invaded staphylococci in endothelial cells. Our data suggest that exocyst complex controls the interaction of recycling endocytic vesicles with phagosomes and this is crucial for the maturation and bactericidal function of PLys in endothelial cells.

  5. Traffic Interchanges

    1. Nucleoli and Stress Granules: Connecting Distant Relatives

      Hicham Mahboubi and Ursula Stochaj

      Article first published online: 22 JUL 2014 | DOI: 10.1111/tra.12191

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      Nucleoli and cytoplasmic stress granules (SGs) are subcellular compartments that modulate the response to endogenous and environmental signals to control cell survival. In our opinion, nucleoli and SGs are functionally linked; they are distant relatives that combine forces when cellular homeostasis is threatened. Given that nucleoli and SGs orchestrate physiological responses that are directly relevant to human health, we propose that several conditions including neurodegeneration, cancer and virus infections will benefit from therapeutic interventions that target simultaneously both compartments.

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