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Cover image for Vol. 16 Issue 5

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

Edited By: Michael S. Marks, Trina A. Schroer, Tom H. Stevens, Sharon A. Tooze

Online ISSN: 1600-0854

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  1. 1 - 12
  1. Original Articles

    1. Lrp1/LDL Receptor Play Critical Roles in Mannose 6-Phosphate-Independent Lysosomal Enzyme Targeting

      Sandra Markmann, Melanie Thelen, Kerstin Cornils, Michaela Schweizer, Nahal Brocke-Ahmadinejad, Thomas Willnow, Joerg Heeren, Volkmar Gieselmann, Thomas Braulke and Katrin Kollmann

      Article first published online: 27 APR 2015 | DOI: 10.1111/tra.12284

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      Stable Isotope Labeling by Amino acids in Cell culture (SILAC)-based quantitative analysis of the lysosomal proteome of mouse fibroblasts allowed the identification of lysosomal enzymes whose localization was unaffected by the lack of mannose 6-phosphate (M6P) targeting signals. Among these, the sorting of cathepsins D and B is independent of sortilin but involves secretion-recapture mechanisms mediated by Lrp1 and LDL receptors.

    2. You have full text access to this OnlineOpen article
      Microtubule Motors Power Plasma Membrane Tubulation in Clathrin-Independent Endocytosis

      Charles A. Day, Nicholas W. Baetz, Courtney A. Copeland, Lewis J. Kraft, Bing Han, Ajit Tiwari, Kimberly R. Drake, Heidi De Luca, Daniel J.-F. Chinnapen, Michael W. Davidson, Randall K. Holmes, Michael G. Jobling, Trina A. Schroer, Wayne I. Lencer and Anne K. Kenworthy

      Article first published online: 27 APR 2015 | DOI: 10.1111/tra.12269

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      How the plasma membrane is bent to accommodate clathrin-independent endocytosis remains uncertain. We studied mechanisms contributing to membrane deformation by imaging the uptake of the cholera toxin B-subunit into surface-attached tubular invaginations that correspond to stalled endocytic intermediates. Unexpectedly, our results suggest microtubules, dynein and dynactin provide an important source of mechanical force that powers tubule elongation, thus defining a novel mechanism for generating membrane curvature in clathrin-independent endocytosis.

    3. Release from Endoplasmic Reticulum Matrix Proteins Controls Cell Surface Transport of MHC Class I Molecules

      Susanne Fritzsche, Esam T. Abualrous, Britta Borchert, Frank Momburg and Sebastian Springer

      Article first published online: 16 APR 2015 | DOI: 10.1111/tra.12279

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      We show that the rate-limiting step of cell surface transport of the major histocompatibility complex class I peptide receptor H-2Db lies prior to endoplasmic reticulum (ER) exit and delays the access of H-2Db to ER exit sites by acting on the folded and peptide-bound lumenal part of the protein. Our data suggest that cell surface transport of class I and other type I transmembrane glycoproteins is governed by the affinity of all folding and maturation states to the proteins of the ER matrix.

    4. Exosomal RNA from Mycobacterium tuberculosis-Infected Cells Is Functional in Recipient Macrophages

      Prachi Pratap Singh, Li Li and Jeffrey Scott Schorey

      Article first published online: 16 APR 2015 | DOI: 10.1111/tra.12278

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      Our study shows a signature of host-derived miRNAs and mRNA transcripts as well as mycobacterial RNA in exosomes derived from Mycobacterium tuberculosis-infected macrophages. The exosomal mRNA was translated upon delivery to recipient cells and was biologically active, suggesting a role in modulation of host response to M. tuberculosis. Mycobacterial transcripts were also detected in extracellular vesicles derived from infected cells pointing to the significance of exosomal RNA in TB diagnostics.

    5. You have full text access to this OnlineOpen article
      A Conserved Di-Basic Motif of Drosophila Crumbs Contributes to Efficient ER Export

      Alexandra Kumichel, Katja Kapp and Elisabeth Knust

      Article first published online: 14 APR 2015 | DOI: 10.1111/tra.12273

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      The evolutionary conserved transmembrane protein Crumbs is required for the maintenance of epithelial polarity. Crumbs protein level at the apical plasma membrane is crucial for polarity, but Crumbs intracellular trafficking is poorly understood. In Drosophila embryos mutant for components of the COPII machinery less Crumbs is delivered to the plasma membrane. We uncovered a di-basic RNKR motif in the cytoplasmic tail of Crumbs, which contributes to efficient ER export in Drosophila Schneider cells and transgenic embryos.

    6. Ultrastructural Morphometry Points to a New Role for LAMTOR2 in Regulating the Endo/Lysosomal System

      Georg F. Vogel, Hannes L. Ebner, Mariana E. G. de Araujo, Thomas Schmiedinger, Oliver Eiter, Haymo Pircher, Karin Gutleben, Barbara Witting, David Teis, Lukas A. Huber and Michael W. Hess

      Article first published online: 14 APR 2015 | DOI: 10.1111/tra.12271

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      The endo/lysosomal LAMTOR/Ragulator complex is essential to control tissue homeostasis via MAPK and mTOR signaling. Quantitative electron microscopy of cryo-fixed LAMTOR2-deficient mouse fibroblasts now indicates that the LAMTOR/Ragulator complex may in addition contribute to the proper formation and turnover of recycling tubules extending from maturing multivesicular endosomes (MVB), thus regulation of membrane traffic from MVB. This further strengthens the key role of the LAMTOR complex in the architecture and function of endo/lysosomes.

    7. You have full text access to this OnlineOpen article
      Surface Trafficking of APP and BACE in Live Cells

      Anna Bauereiss, Oliver Welzel, Jasmin Jung, Simon Grosse-Holz, Natalia Lelental, Piotr Lewczuk, Eva M. Wenzel, Johannes Kornhuber and Teja W. Groemer

      Article first published online: 14 APR 2015 | DOI: 10.1111/tra.12270

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      In this study, we demonstrated that total internal reflection fluorescence (TIRF) imaging of pHluorin-amyloid-β precursor protein (pHAPP) and pHluorin-β-site APP-cleaving enzyme (pHBACE) can be used to elucidate the exocytosis of amyloid-β precursor protein (APP) and β-site APP-cleaving enzyme (BACE). We found that APP and BACE exhibit distinct fusion kinetics and surface trafficking. Furthermore, BACE and the transferrin receptor (TfR) are transported and released together, and the fusion frequencies of APP and BACE are distinct and can be pharmacologically altered using methyl-β-cyclodextrin (MβCD).

  2. Toolbox

    1. High-Resolution Membrane Capacitance Measurements for Studying Endocytosis and Exocytosis in Yeast

      Lucia Carrillo, Bayram Cucu, Vera Bandmann, Ulrike Homann, Brigitte Hertel, Stefan Hillmer, Gerhard Thiel and Adam Bertl

      Article first published online: 9 APR 2015 | DOI: 10.1111/tra.12275

      With patch-clamp recording, we detect in yeast protoplasts individual exo- and endocytotic events as discrete steps in membrane capacitance. The high-resolution data show that exo- and endocytotic vesicles undergo, similar to other eukaryotes, permanent and transient fusion/fission with a bias for transient events. The electrical data are a good representation of the membrane dynamics of a growing yeast cell.

  3. Original Articles

    1. Phenothiazine-Derived Antipsychotic Drugs Inhibit Dynamin and Clathrin-Mediated Endocytosis

      James A. Daniel, Ngoc Chau, Mohammed K. Abdel-Hamid, Lingbo Hu, Lisa von Kleist, Ainslie Whiting, Sai Krishnan, Peter Maamary, Shannon R. Joseph, Fiona Simpson, Volker Haucke, Adam McCluskey and Phillip J. Robinson

      Article first published online: 9 APR 2015 | DOI: 10.1111/tra.12272

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      We report that all clinically approved phenothiazine antipsychotic drugs (APDs), including chlorpromazine, are lipid-competitive inhibitors of dynamin I and II in vitro and of clathrin-mediated endocytosis (CME) in cells. Some, but not all non-phenothiazine APDs also inhibit dynamin and CME. Thus dynamin inhibition is common to all phenothiazine-based APDs, but to not all non-phenothiazine APDs. Chlorpromazine does not inhibit clathrin or AP-2 recruitment to the membrane, as previously proposed, indicating that dynamin inhibition is their likely cellular target for CME inhibition.

    2. CD14, TLR4 and TRAM Show Different Trafficking Dynamics During LPS Stimulation

      Dionne C.G. Klein, Astrid Skjesol, Esther D. Kers-Rebel, Tatyana Sherstova, Bjørnar Sporsheim, Kjartan W. Egeberg, Bjørn T. Stokke, Terje Espevik and Harald Husebye

      Article first published online: 24 MAR 2015 | DOI: 10.1111/tra.12274

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      We have investigated the location and mobility of toll-like receptor 4 (TLR4) and its adaptor TRAM during lipopolysaccharide (LPS)-induced signaling and how RAB11A regulates TRAM trafficking. We show that LPS induces an immobile fraction of TLR4 in punctate structures containing CD14/LPS and clathrin. RAB11A drives TRAM into the endosomal recycling compartment (ERC) and onto early sorting endosomes. TRAM is recovered more from sorting endosomes than CD14/LPS. Our data suggest that RAB11A regulates LPS-induced interferon-β (IFN-β) production through its ability to transport TRAM form the Golgi to the ERC and further onto sorting endosomes.

    3. Lysosomal Targeting of Cystinosin Requires AP-3

      Zuzanna Andrzejewska, Nathalie Névo, Lucie Thomas, Anne Bailleux, Véronique Chauvet, Alexandre Benmerah and Corinne Antignac

      Article first published online: 24 MAR 2015 | DOI: 10.1111/tra.12277

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      Cystinosin, a seven transmembrane (TM) lysosomal cystine transporter defective in the autosomal recessive lysosomal storage disorder, has been shown to be targeted to lysosomes via two sorting signals: a tyrosine-based motif in its C-terminal (GYDQL) and a non-classical motif in its fifth inter-TM loop. Here, we show that the tyrosine-based motif specifically interacts with AP-3 complex and functions as a ‘strong’ AP-3 motif responsible for direct intracellular targeting of cystinosin and the CD63-cystinosin chimera (CD63-GYDQL) to late endosomes/lysosomes.

  4. Corrigendum

    1. You have free access to this content
      The RAB5-GEF Function of RIN1 Regulates Multiple Steps During Listeria monocytogenes Infection

      Kavitha Balaji, Christopher T. French, Jeff F. Miller and John Colicelli

      Article first published online: 24 FEB 2015 | DOI: 10.1111/tra.12268

      This article corrects:

      The RAB5-GEF Function of RIN1 Regulates Multiple Steps During Listeria monocytogenes Infection

      Vol. 15, Issue 11, 1206–1218, Article first published online: 4 SEP 2014

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