Traffic

Cover image for Vol. 15 Issue 11

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

Online ISSN: 1600-0854

Highlights

  • ORIGINAL ARTICLE: Vmp1 Regulates PtdIns3P Signaling During Autophagosome Formation in Dictyostelium discoideum

    ORIGINAL ARTICLE: Vmp1 Regulates PtdIns3P Signaling During Autophagosome Formation in Dictyostelium discoideum

    The pattern of GFP-Atg18 in the vmp1− mutant suggests a defect in an early stage of autophagosome formation. A) Dictyostelium cells expressing GFP-Atg18 and Vmp1-RFP were imaged by time-lapse confocal fluorescence to show colocalization. B) Wild-type and mutant cells expressing the autophagosome marker GFP-Atg18 were analysed by confocal microscopy during growth in HL5 and starvation in PDF for 30 min. Maximum intensity projections of 10 z-optical sections are shown. B) Sequential z-sections of the cell labeled by and asterisk in panel B showing an abnormal ring-like feature. Bars: 10 µm.

  • ORIGINAL ARTICLE: Targeting of Viral Capsids to Nuclear Pores in a Cell-Free Reconstitution System

    ORIGINAL ARTICLE: Targeting of Viral Capsids to Nuclear Pores in a Cell-Free Reconstitution System

    Targeting of HSV1 capsids to NPCs on Xenopus nuclei in vitro. A–C). Functional (A), Imp β45–462 (B) or BAPTA (C) nuclei were reconstituted in vitro. Completion of nuclear assembly was monitored by epi-fluorescence microscopy by DNA staining with Hoechst 33258 (i and iii), NPC staining with fluorescently labeled mAb414 (ii) and the transport capacity of the assembled NPCs was confirmed by the nuclear import of TRITC-NLS-BSA (iv). Scale bar: 10 µm. D,E) Binding of viral 0.5 m KCl capsids extracted from extracellular particles of HSV1-GFPVP26 (ii and iii, green, white arrowheads) to Xenopus nuclei was analyzed using confocal fluorescence laser scanning microscopy and a serial z-slicing of 0.37 µm optical sections. A surface view (D) and one midsection (E) of a representative nucleus are shown. NPCs were labeled with Imp β45–462-TRITC (i and iii, red). Scale bar: 10 µm. F,G) HSV1-GFPVP26 viral 0.5 m capsids were incubated with Xenopus nuclei harboring functional NPCs for 45 min, labeled with a polyclonal rabbit antiserum raised against intact HSV1 capsids (Remus, bleed V) and colloidal gold with a diameter of 12 nm coated with anti-rabbit antibodies. The specimens were analyzed by field emission scanning electron microscopy. F) 3D surface topography of reconstituted nuclei from different views in the in-lens images (i–iii). The same areas were imaged through a backscatter electron detector revealing the positions of gold-conjugated antibodies (Gi–iii; inverted color mode). HSV1 capsids (white arrowheads) are bound to the cytoplasmic face of the NPCs (white arrows). Scale bar: 100 nm.

  • ORIGINAL ARTICLE : Arabinogalactan Glycosyltransferases Target to a Unique Subcellular Compartment That May Function in Unconventional Secretion in Plants

    ORIGINAL ARTICLE : Arabinogalactan Glycosyltransferases Target to a Unique Subcellular Compartment That May Function in Unconventional Secretion in Plants

    The subcellular localization ofAtGlcAT14A-mCer3. AtGlcAT14A-mCer3 (A–I, green) was coexpressed with GnTI-mRFP (A–C, magenta), GMII-mRFP (D–F, magenta) and ST-YFP (G–I, magenta) in N. benthamiana leaves. AtGlcAT14A-mCer3 colocalized with GnTI-mRFP, GMII-mRFP and ST-YFP in the Golgi compartments (overlapping signals, A–I) but was also found alone in the small compartments, indicated by arrowheads. Scale bars = 10 µm.

  • ORIGINAL ARTICLE: Nucleoli and Stress Granules: Connecting Distant Relatives

    ORIGINAL ARTICLE: Nucleoli and Stress Granules: Connecting Distant Relatives

    Localization of nucleolar andSGproteins in stressed mammalian cells. HeLa cells were treated with sodium arsenite, a compound that induces oxidative stress, and proteins were detected by immunofluorescence staining. Nucleostemin (magenta) is a marker for nucleoli, whereas G3BP1 (yellow) associates with SGs. Note that the RNA-binding protein hnRNP K (green) is present in both compartments. Surface rendering was performed on confocal image stacks; size bar is 5 µm (A). Magnified views of a nucleolus (B), the nucleolar interior (C) or one of the SGs (D).

  • ORIGINAL ARTICLE: Immuno- and Correlative Light Microscopy-Electron Tomography Methods for 3D Protein Localization in Yeast

    ORIGINAL ARTICLE: Immuno‐ and Correlative Light Microscopy‐Electron Tomography Methods for 3D Protein Localization in Yeast

    3D reconstruction of a yeast mitochondrion in consecutive serial sections. Three consecutive serial sections (each 250 nm thick) of embedded FRY344 cells mixed with human blood cells (see Materials and Methods), were cut and used for immunogold labelling using the anti-HA antibody and 10 nm gold particle-conjugated protein A. In addition 15 nm fiducial gold particles were applied. A–C) Cells of interest were imaged at low magnification in three consecutive sections ((A) section 1; (B) section 2; (C) section 3), using a Tecnai20 electron microscope. Blood cells surrounding the yeast cell of interest are pointed-out with a square, a circle and a star, and where used to localize the yeast cell in the consecutive sections. D) Tomographic slice of the second serial section (see (B)) showing organelle structures of the yeast cell. E) Example of contouring different compartments in a tomographic slice of the yeast cell. F and G) Model of the entire yeast cell through the three sections. H) Rotation of about 180° along the x-axis of the 3D model. I) Enlargement of the ER/mitochondria area displaying the Idh1-3XHA immunogold labelling (red dots). The entire tomogram and model are shown in Movie S3. ER, endoplasmic reticulum (yellow); G, Golgi (light blue); M, mitochondrion (dark red); PM, plasma membrane (white); V, vacuole (light grey with transparency); N, nucleus (orange). Please note that only part of the ER was drawn to allow distinctively seeing the other cell organelles. Scale bar, 3.5 µm (A–C), 700 nm (D–F).

  • ORIGINAL ARTICLE: Vmp1 Regulates PtdIns3P Signaling During Autophagosome Formation in Dictyostelium discoideum
  • ORIGINAL ARTICLE: Targeting of Viral Capsids to Nuclear Pores in a Cell-Free Reconstitution System
  • ORIGINAL ARTICLE : Arabinogalactan Glycosyltransferases Target to a Unique Subcellular Compartment That May Function in Unconventional Secretion in Plants
  • ORIGINAL ARTICLE: Nucleoli and Stress Granules: Connecting Distant Relatives
  • ORIGINAL ARTICLE: Immuno‐ and Correlative Light Microscopy‐Electron Tomography Methods for 3D Protein Localization in Yeast

Recently Published Issues

See all

Video Highlight

July 2014 Video Highlight

"Rab5 and Ndfip1 Are Involved in Pten Ubiquitination and Nuclear Trafficking" - Yi et al

Top Cited articles of 2013

Endogenous GSK-3/Shaggy Regulates Bidirectional Axonal Transport of the Amyloid Precursor Protein
Carole Weaver, Christina Leidel, Lukasz Szpankowski, Nicole M. Farley, George T. Shubeita and Lawrence S. B. Goldstein

Disulfide Bond Formation: Sulfhydryl Oxidase ALR Controls Mitochondrial Biogenesis of Human MIA40
Malgorzata E. Sztolsztener, Anita Brewinska, Bernard Guiard and Agnieszka Chacinska

Highlight

Find out more

Rab5 and Ndfip1 Are Involved in Pten Ubiquitination and Nuclear Trafficking
Yijia Li, Ley-Hian Low, Ulrich Putz, Choo-Peng Goh, Seong-Seng Tan and Jason Howitt

Depending on its localization, Pten (the central antagonist of PI3K signaling in the cytoplasm) is involved in many diverse cellular functions including controlling mitosis and DNA repair, cellular homeostasis, cell migration and/or cell proliferation. Balancing the cellular distribution of Pten is crucial to the function of the cell. Li and colleagues provide evidence that sorting of Pten to various organelles occurs in endosomes. Using bimolecular fluorescence complementation and dominant negative Rab5, they demonstrate that Rab5 and the E3 ligase adaptor protein Ndfip1 work together in to ubiquitinate Pten, which is required for its trafficking to the nucleus.


Traf

SEARCH

SEARCH BY CITATION