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Cover image for Vol. 15 Issue 6

June 2014

Volume 15, Issue 6

Pages i–i, 583–726

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  2. Original Articles

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      Ultrafast Diffusion of a Fluorescent Cholesterol Analog in Compartmentalized Plasma Membranes (pages 583–612)

      Nao Hiramoto-Yamaki, Kenji A. K. Tanaka, Kenichi G. N. Suzuki, Koichiro M. Hirosawa, Manami S. H. Miyahara, Ziya Kalay, Koichiro Tanaka, Rinshi S. Kasai, Akihiro Kusumi and Takahiro K. Fujiwara

      Version of Record online: 11 MAR 2014 | DOI: 10.1111/tra.12163

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      A fluorescent cholesterol analogue, Bodipy488-conjugated cholesterol molecule (Bdp-Chol), exhibited diffusion at the fastest rate ever found for any membrane molecules in the plasma membrane, with a median diffusion coefficient of 3.4 µm2/second, ∼10× greater than that of phospholipids, despite Bdp-Chol's probable association with raft domains. Bdp-Chol and phospholipids diffused at comparable rates in the absence of actin membrane-skeleton, indicating that Bdp-Chol hops across the actin-fence-pickets' compartment boundaries ∼10× faster than phospholipids. Raft domains coexist with membrane-skeleton-induced compartments and are contained within them.

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      Sec16 Determines the Size and Functioning of the Golgi in the Protist Parasite, Trypanosoma brucei (pages 613–629)

      Marco Sealey-Cardona, Katy Schmidt, Lars Demmel, Tatjana Hirschmugl, Tanja Gesell, Gang Dong and Graham Warren

      Version of Record online: 4 APR 2014 | DOI: 10.1111/tra.12170

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      Changes in TbSec16 levels affect the size of the endoplasmic reticulum exit site (ERES) and Golgi (TbGRASP). Depletion of TbSec16 (top panel) reduces the size of the ERES and in turn the Golgi, decreasing the flux of anterograde cargo and the growth rate. In contrast, an increase in the levels of TbSec16 (bottom panel) generates a bigger ERES and Golgi (TbGRASP) without any significant impact on the cargo flux or growth rate. This suggests that the size of the ERES and the Golgi is set for optimal growth and cargo flux in the protist parasite, Trypanosoma brucei.

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      Distinct Sets of Rab6 Effectors Contribute to ZW10- and COG-Dependent Golgi Homeostasis (pages 630–647)

      Waqar Majeed, Shijie Liu and Brian Storrie

      Version of Record online: 11 APR 2014 | DOI: 10.1111/tra.12167

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      ZW10 and COG3 are key proteins in separate, retrograde tether protein complexes important to the organization of the mammalian Golgi apparatus. Knockdown of either results in the fragmentation and dispersal of the juxtanuclear Golgi ribbon. We show that a small and partially overlapping set of motor protein-linked, Rab6 effectors are important to the two pathways. Myosin II and Kif20A, selectively contribute to the ZW10 pathway while Bicaudal D1/D2, like Rab6 itself, is important to both pathways. The order in which proteins act is inferred from experimental data Isee Discussion).

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      Polarised Clathrin-Mediated Endocytosis of EGFR During Chemotactic Invasion (pages 648–664)

      Laura Jane Mutch, Jake Davey Howden, Emma Poppy Louise Jenner, Natalie Sarah Poulter and Joshua Zachary Rappoport

      Version of Record online: 20 MAR 2014 | DOI: 10.1111/tra.12165

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      Using a novel assay for chemotactic invasion we were able to show that clathrin-mediated endocytosis, but not caveolar endocytosis, is necessary for migration of MDA-MB-231 cells towards EGF. We show that the clathrin-mediated endocytosis pathway is involved in internalisation of EGFR and is polarised towards the front of migrating cells. We also show that clathrin-mediated endocytosis plays no role in the disassembly of focal adhesions in our experimental system.

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      Photoactivation Approaches Reveal a Role for Rab11 in FGFR4 Recycling and Signalling (pages 665–683)

      Ellen M. Haugsten, Andreas Brech, Knut Liestøl, Jim C. Norman and Jørgen Wesche

      Version of Record online: 31 MAR 2014 | DOI: 10.1111/tra.12168

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      Direct visualization of FGFR4 recycling using a novel photoactivatable probe. In Rab11b-depleted cells, FGFR4 is not delivered to lysosomes, but accumulates in perinuclear vesicles, leading to increased cellular receptor levels and altered FGFR4 signalling.

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      A 14-3-3 Mode-1 Binding Motif Initiates Gap Junction Internalization During Acute Cardiac Ischemia (pages 684–699)

      James W. Smyth, Shan-Shan Zhang, Jose M. Sanchez, Samy Lamouille, Jacob M. Vogan, Geoffrey G. Hesketh, TingTing Hong, Gordon F. Tomaselli and Robin M. Shaw

      Version of Record online: 9 APR 2014 | DOI: 10.1111/tra.12169

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      Cardiac ischemia rapidly leads to alterations in phosphorylation status and altered localization of connexin 43 (Cx43) gap junctions, resulting in ventricular arrhythmias. In this study, we find that phosphorylation-mediated activation of a 14-3-3 binding motif at Ser373 within the Cx43 C-terminus is a gatekeeper event necessary for further phosphorylation and ubiquitination of Cx43, leading to endocytosis of gap junctions. Regulation of Cx43Ser373 phosphorylation and Cx43/14-3-3 complexing could be a therapeutic strategy to restore gap junction coupling in ischemic disease.

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      The CryoCapsule: Simplifying Correlative Light to Electron Microscopy (pages 700–716)

      Xavier Heiligenstein, Jérôme Heiligenstein, Cédric Delevoye, Ilse Hurbain, Sabine Bardin, Perrine Paul-Gilloteaux, Lucie Sengmanivong, Gilles Régnier, Jean Salamero, Claude Antony and Graca Raposo

      Version of Record online: 24 MAR 2014 | DOI: 10.1111/tra.12164

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      Correlating complementary multiple scale images of the same object is a straightforward means to decipher biological processes. Light microscopy and electron microscopy are the most commonly used imaging techniques, yet despite their complementarity, the experimental procedures available to correlate them are technically complex. We designed and manufactured a new device adapted to many biological specimens, the CryoCapsule, that simplifies the multiple sample preparation steps, which at present separate live cell fluorescence imaging from contextual high-resolution electron microscopy.

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