DOCSLIB.ORG

RAB11-Mediated Trafficking and Human Cancers: an Updated Review

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

biology Review RAB11-Mediated Trafficking and Human Cancers: An Updated Review Elsi Ferro 1,2, Carla Bosia 1,2 and Carlo C. Campa 1,2,* 1 Department of Applied Science and Technology, Politecnico di Torino, 24 Corso Duca degli Abruzzi, 10129 Turin, Italy; [email protected] (E.F.); [email protected] (C.B.) 2 Italian Institute for Genomic Medicine, c/o IRCCS, Str. Prov. le 142, km 3.95, 10060 Candiolo, Italy * Correspondence: [email protected] Simple Summary: The small GTPase RAB11 is a master regulator of both vesicular trafficking and membrane dynamic defining the surface proteome of cellular membranes. As a consequence, the alteration of RAB11 activity induces changes in both the sensory and the transduction apparatuses of cancer cells leading to tumor progression and invasion. Here, we show that this strictly depends on RAB110s ability to control the sorting of signaling receptors from endosomes. Therefore, RAB11 is a potential therapeutic target over which to develop future therapies aimed at dampening the acquisition of aggressive traits by cancer cells. Abstract: Many disorders block and subvert basic cellular processes in order to boost their pro- gression. One protein family that is prone to be altered in human cancers is the small GTPase RAB11 family, the master regulator of vesicular trafficking. RAB11 isoforms function as membrane organizers connecting the transport of cargoes towards the plasma membrane with the assembly of autophagic precursors and the generation of cellular protrusions. These processes dramatically impact normal cell physiology and their alteration significantly affects the survival, progression and metastatization as well as the accumulation of toxic materials of cancer cells. In this review, we dis- cuss biological mechanisms ensuring cargo recognition and sorting through a RAB11-dependent pathway, a prerequisite to understand the effect of RAB11 alterations in human cancers. Citation: Ferro, E.; Bosia, C.; Campa, Keywords: RAB11; RAB25; endosome; sorting; cancer C.C. RAB11-Mediated Trafficking and Human Cancers: An Updated Review. Biology 2021, 10, 26. https://doi.org/ 10.3390/biology10010026 1. Introduction Received: 15 November 2020 To manage pathogenic alterations induced by genetic/epigenetic/transcriptomic Accepted: 30 December 2020 changes, cells evolved various strategies to exploit cellular machinery and guarantee their Published: 4 January 2021 own survival. Specific genetic alterations modify key elements controlling the molecular composition of the cell surface, the assembly of internal organelles and the efficacy of Publisher’s Note: MDPI stays neu- intracellular and extracellular transmission. RAB GTPases are key players in membrane tral with regard to jurisdictional clai- transport events and are fundamental to ensure the efficacy of intracellular logistics [1–3]. ms in published maps and institutio- RAB GTPases are members of the Ras superfamily of monomeric G proteins, a large nal affiliations. protein family which includes among its members RAS, RHO–RAC, ARF and RAN as well as the more recently characterized RHEB, RAD and RIT subfamilies [4]. Similarly to other small GTPases, Rab proteins are molecular switches that cycle between an active and an inactive state through the association and the subsequent hydrolysis of guanosine-50- Copyright: © 2021 by the authors. Li- censee MDPI, Basel, Switzerland. triphosphate (GTP). In particular, association with GTP is stimulated by Guanine nucleotide This article is an open access article exchange factors (GEFs), whereas hydrolysis of GTP to GDP is promoted by GTPase- distributed under the terms and con- activating proteins (GAPs) [5]. This activation cycle is pivotal for the binding of multiple ditions of the Creative Commons At- effector proteins which mediate the delivery of cellular material to various intracellular tribution (CC BY) license (https:// compartments such as—the plasma membrane, endosomes, autophagosomes, leading creativecommons.org/licenses/by/ edge of migrating cells, midbodies, primary cilium and centrioles [6]. As a matter of fact, 4.0/). alterations of RAB-mediated trafficking are pivotal in various aspects of both progression Biology 2021, 10, 26. https://doi.org/10.3390/biology10010026 https://www.mdpi.com/journal/biology Biology 2021, 10, 26 2 of 15 Biology 2021, 10, x 2 of 15 and tumorgenicity of cancer cells including the sustainment of proliferative signaling, the evasion of growth suppression, the induction of receptor recycling, the activation of receptor invasion recycling, and metastasis the activation as well of as invasi the reprogrammingon and metastasis of as tumor well as metabolism the reprogramming and the ofevasion tumor ofmetabolism immune destruction and the evasio [7,8n]. of immune destruction [7,8]. 2. Different Different RAB11 RAB11 Isoforms Isoforms fo forr Distinct Distinct Cellular Cellular Functions Functions OneOne prominent prominent Rab-mediated Rab-mediated transport transport pathway pathway that that cancer cancer cells cells exploit exploit to adapt to adapt their internaltheir internal states statesto fluctuations to fluctuations of both of their both gene their expression gene expression and microenvironmental and microenvironmental status is thestatus RAB11-mediated is the RAB11-mediated trafficking trafficking pathway. pathway. This transport This transport route relies route on relies members on members of the RAB11of the RAB11 protein protein family familywhich control which controlthe delivery the delivery of both ofproteins both proteins and lipids and toward lipids towardseveral organellesseveral organelles and therefore and therefore are involved are involvedin many cellular in many processes cellular (Figure processes 1A,B). (Figure 1A,B). Figure 1. Overview of the main cellular processes and traffickingtrafficking pathways regulated by RAB11 isoforms. (A) Schematic representation of of cellular cellular processes processes controlled controlled by by RAB11. RAB11. Blac Blackk lines lines represent represent cells’ cells’ contour, contour, green green arrows arrows represent represent di- rections of RAB11-mediated trafficking pathways. (B) Schematic representation of RAB11 isoform localization in both directions of RAB11-mediated trafficking pathways. (B) Schematic representation of RAB11 isoform localization in both endocytic (left panel) and exocytic pathways (right panel). Green circles represent RAB11-positive compartments, yellow, endocytic (left panel) and exocytic pathways (right panel). Green circles represent RAB11-positive compartments, yellow, blue, red lines represent RAB11A, RAB11B and RAB25, respectively. blue, red lines represent RAB11A, RAB11B and RAB25, respectively. The RAB11 protein family includes three different isoforms named RAB11A, RAB11B The RAB11 protein family includes three different isoforms named RAB11A, RAB11B and and RAB25, each one encoded by a different gene located in a distinct chromosome (Table RAB25, each one encoded by a different gene located in a distinct chromosome 1). All RAB11 isoforms are widely distributed in human tissues. In particular, while (Table1). All RAB11 isoforms are widely distributed in human tissues. In particular, RAB11A is ubiquitously expressed, both RAB11B and RAB25 are enriched in specific or- while RAB11A is ubiquitously expressed, both RAB11B and RAB25 are enriched in specific gansorgans (Table (Table 1)1 [9].)[ 9 Notably,]. Notably, RAB11 RAB11 proteins proteins di differffer in in their their percentage percentage of of sequence sequence identity. identity. While RAB11ARAB11A andand RAB11B RAB11B share share 89% 89% of sequenceof sequence similarity, similarity, the identity the identity between between RAB25 RAB25with either with RAB11A either RAB11A or RAB11B or RAB11B is lower is than lower 70% than (Table 70%1 )[(Table10]. 1) [10]. Although minimal, such differences in protein sequence identity significantly impact Table 1. Structure of RAB11 gene isoforms. both protein structure and specificity of effector binding, providing an explanation for the different roles ofChromosome RAB11 isoforms Number in cells of [11 ,12Number]. All RAB11 of proteinsPercentage present of severalTissue post- Gene Name translational modifications,Position includingCoding Exon prenylation, Amino phosphorylationAcids Identity and ubiquitination.Expression In particular, while prenylation at RAB11 C-termini allows Rab100% membrane association, RAB11A 15q22.31 5 216 Ubiquitous both phosphorylation and ubiquitination modulate either activity(with orRAB11A) degradation of RAB11 isoforms, respectively [9]. Wide, RAB11A is the first discovered and best characterized member89% of the RAB11enriched subfamily. in RAB11B 19p13.2 5 218 Homozygous depletion of Rab11a in mice is embryo lethal;(with whereas RAB11A) the loss brain of RAB11A testis, protein in neurons does not significantly impact both brain development and functionality.heart In contrast, depletion of Rab11a in mice intestine increases intracellular accumulationWide, of 61% and 66% enriched in lung apicalRAB25 proteins and it1q22 induces shortening5 of microvilli213 [13,14].(with RAB11A kidney, gastric and RAB11B) tract Biology 2021, 10, 26 3 of 15 Table 1. Structure of RAB11 gene isoforms. Number of Chromosome Number of Percentage Tissue Gene Name Amino Position Coding Exon of Identity Expression Acids 100% RAB11A 15q22.31 5 216 (with Ubiquitous RAB11A) Wide, 89% enriched in RAB11B 19p13.2 5 218 (with brain testis, RAB11A) heart 61% and 66% Wide, (with enriched in RAB25
Recommended publications
  • Supplementary Data

    Supplementary Data

    Figure 2S 4 7 A - C 080125 CSCs 080418 CSCs - + IFN-a 48 h + IFN-a 48 h + IFN-a 72 h 6 + IFN-a 72 h 3 5 MRFI 4 2 3 2 1 1 0 0 MHC I MHC II MICA MICB ULBP-1 ULBP-2 ULBP-3 ULBP-4 MHC I MHC II MICA MICB ULBP-1 ULBP-2 ULBP-3 ULBP-4 7 B 13 080125 FBS - D 080418 FBS - + IFN-a 48 h 12 + IFN-a 48 h + IFN-a 72 h + IFN-a 72 h 6 080125 FBS 11 10 5 9 8 4 7 6 3 MRFI 5 4 2 3 2 1 1 0 0 MHC I MHC II MICA MICB ULBP-1 ULBP-2 ULBP-3 ULBP-4 MHC I MHC II MICA MICB ULBP-1 ULBP-2 ULBP-3 ULBP-4 Molecule Molecule FIGURE 4S FIGURE 5S Panel A Panel B FIGURE 6S A B C D Supplemental Results Table 1S. Modulation by IFN-α of APM in GBM CSC and FBS tumor cell lines. Molecule * Cell line IFN-α‡ HLA β2-m# HLA LMP TAP1 TAP2 class II A A HC§ 2 7 10 080125 CSCs - 1∞ (1) 3 (65) 2 (91) 1 (2) 6 (47) 2 (61) 1 (3) 1 (2) 1 (3) + 2 (81) 11 (80) 13 (99) 1 (3) 8 (88) 4 (91) 1 (2) 1 (3) 2 (68) 080125 FBS - 2 (81) 4 (63) 4 (83) 1 (3) 6 (80) 3 (67) 2 (86) 1 (3) 2 (75) + 2 (99) 14 (90) 7 (97) 5 (75) 7 (100) 6 (98) 2 (90) 1 (4) 3 (87) 080418 CSCs - 2 (51) 1 (1) 1 (3) 2 (47) 2 (83) 2 (54) 1 (4) 1 (2) 1 (3) + 2 (81) 3 (76) 5 (75) 2 (50) 2 (83) 3 (71) 1 (3) 2 (87) 1 (2) 080418 FBS - 1 (3) 3 (70) 2 (88) 1 (4) 3 (87) 2 (76) 1 (3) 1 (3) 1 (2) + 2 (78) 7 (98) 5 (99) 2 (94) 5 (100) 3 (100) 1 (4) 2 (100) 1 (2) 070104 CSCs - 1 (2) 1 (3) 1 (3) 2 (78) 1 (3) 1 (2) 1 (3) 1 (3) 1 (2) + 2 (98) 8 (100) 10 (88) 4 (89) 3 (98) 3 (94) 1 (4) 2 (86) 2 (79) * expression of APM molecules was evaluated by intracellular staining and cytofluorimetric analysis; ‡ cells were treatead or not (+/-) for 72 h with 1000 IU/ml of IFN-α; # β-2 microglobulin; § β-2 microglobulin-free HLA-A heavy chain; ∞ values are indicated as ratio between the mean of fluorescence intensity of cells stained with the selected mAb and that of the negative control; bold values indicate significant MRFI (≥ 2).
  • Anti-Rab11 Antibody (ARG41900)

    Anti-Rab11 Antibody (ARG41900)

    Product datasheet [email protected] ARG41900 Package: 100 μg anti-Rab11 antibody Store at: -20°C Summary Product Description Goat Polyclonal antibody recognizes Rab11 Tested Reactivity Hu, Ms, Rat, Dog, Mk Tested Application IHC-Fr, IHC-P, WB Host Goat Clonality Polyclonal Isotype IgG Target Name Rab11 Antigen Species Mouse Immunogen Purified recombinant peptides within aa. 110 to the C-terminus of Mouse Rab11a, Rab11b and Rab11c (Rab25). Conjugation Un-conjugated Alternate Names RAB11A: Rab-11; Ras-related protein Rab-11A; YL8 RAB11B: GTP-binding protein YPT3; H-YPT3; Ras-related protein Rab-11B RAB25: RAB11C; CATX-8; Ras-related protein Rab-25 Application Instructions Application table Application Dilution IHC-Fr 1:100 - 1:400 IHC-P 1:100 - 1:400 WB 1:250 - 1:2000 Application Note IHC-P: Antigen Retrieval: Heat mediation was recommended. * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. Positive Control Hepa cell lysate Calculated Mw 24 kDa Observed Size ~ 26 kDa Properties Form Liquid Purification Affinity purification with immunogen. Buffer PBS, 0.05% Sodium azide and 20% Glycerol. Preservative 0.05% Sodium azide www.arigobio.com 1/3 Stabilizer 20% Glycerol Concentration 3 mg/ml Storage instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use. Note For laboratory research only, not for drug, diagnostic or other use.
  • In Silico Prediction of High-Resolution Hi-C Interaction Matrices

    In Silico Prediction of High-Resolution Hi-C Interaction Matrices

    ARTICLE https://doi.org/10.1038/s41467-019-13423-8 OPEN In silico prediction of high-resolution Hi-C interaction matrices Shilu Zhang1, Deborah Chasman 1, Sara Knaack1 & Sushmita Roy1,2* The three-dimensional (3D) organization of the genome plays an important role in gene regulation bringing distal sequence elements in 3D proximity to genes hundreds of kilobases away. Hi-C is a powerful genome-wide technique to study 3D genome organization. Owing to 1234567890():,; experimental costs, high resolution Hi-C datasets are limited to a few cell lines. Computa- tional prediction of Hi-C counts can offer a scalable and inexpensive approach to examine 3D genome organization across multiple cellular contexts. Here we present HiC-Reg, an approach to predict contact counts from one-dimensional regulatory signals. HiC-Reg pre- dictions identify topologically associating domains and significant interactions that are enri- ched for CCCTC-binding factor (CTCF) bidirectional motifs and interactions identified from complementary sources. CTCF and chromatin marks, especially repressive and elongation marks, are most important for HiC-Reg’s predictive performance. Taken together, HiC-Reg provides a powerful framework to generate high-resolution profiles of contact counts that can be used to study individual locus level interactions and higher-order organizational units of the genome. 1 Wisconsin Institute for Discovery, 330 North Orchard Street, Madison, WI 53715, USA. 2 Department of Biostatistics and Medical Informatics, University of Wisconsin-Madison, Madison, WI 53715, USA. *email: [email protected] NATURE COMMUNICATIONS | (2019) 10:5449 | https://doi.org/10.1038/s41467-019-13423-8 | www.nature.com/naturecommunications 1 ARTICLE NATURE COMMUNICATIONS | https://doi.org/10.1038/s41467-019-13423-8 he three-dimensional (3D) organization of the genome has Results Temerged as an important component of the gene regulation HiC-Reg for predicting contact count using Random Forests.
  • Ras Gtpase Chemi ELISA Kit Catalog No

    Ras Gtpase Chemi ELISA Kit Catalog No

    Ras GTPase Chemi ELISA Kit Catalog No. 52097 (Version B3) Active Motif North America 1914 Palomar Oaks Way, Suite 150 Carlsbad, California 92008, USA Toll free: 877 222 9543 Telephone: 760 431 1263 Fax: 760 431 1351 Active Motif Europe Waterloo Atrium Drève Richelle 167 – boîte 4 BE-1410 Waterloo, Belgium UK Free Phone: 0800 169 31 47 France Free Phone: 0800 90 99 79 Germany Free Phone: 0800 181 99 10 Telephone: +32 (0)2 653 0001 Fax: +32 (0)2 653 0050 Active Motif Japan Azuma Bldg, 7th Floor 2-21 Ageba-Cho, Shinjuku-Ku Tokyo, 162-0824, Japan Telephone: +81 3 5225 3638 Fax: +81 3 5261 8733 Active Motif China 787 Kangqiao Road Building 10, Suite 202, Pudong District Shanghai, 201315, China Telephone: (86)-21-20926090 Hotline: 400-018-8123 Copyright 2021 Active Motif, Inc. www.activemotif.com Information in this manual is subject to change without notice and does not constitute a commit- ment on the part of Active Motif, Inc. It is supplied on an “as is” basis without any warranty of any kind, either explicit or implied. Information may be changed or updated in this manual at any time. This documentation may not be copied, transferred, reproduced, disclosed, or duplicated, in whole or in part, without the prior written consent of Active Motif, Inc. This documentation is proprietary information and protected by the copyright laws of the United States and interna- tional treaties. The manufacturer of this documentation is Active Motif, Inc. © 2021 Active Motif, Inc., 1914 Palomar Oaks Way, Suite 150; Carlsbad, CA 92008.
  • Myosin Vb Mediates Cu+ Export in Polarized Hepatocytes Arnab Gupta1,*,‡, Michael J

    Myosin Vb Mediates Cu+ Export in Polarized Hepatocytes Arnab Gupta1,*,‡, Michael J

    © 2016. Published by The Company of Biologists Ltd | Journal of Cell Science (2016) 129, 1179-1189 doi:10.1242/jcs.175307 RESEARCH ARTICLE Myosin Vb mediates Cu+ export in polarized hepatocytes Arnab Gupta1,*,‡, Michael J. Schell1, Ashima Bhattacharjee2, Svetlana Lutsenko2 and Ann L. Hubbard1 ABSTRACT myosin Vc (MYO5C; hereafter referred to as MyoVa and The cellular machinery responsible for Cu+-stimulated delivery of the MyoVc, respectively) are not highly expressed. Mutations in the MYO5B Wilson-disease-associated protein ATP7B to the apical domain of human gene lead to microvillus inclusion disease hepatocytes is poorly understood. We demonstrate that myosin Vb (MVID), in which the filamentous actin (F-actin)-rich apical regulates the Cu+-stimulated delivery of ATP7B to the apical domain microvilli of enterocytes are absent, with concomitantly of polarized hepatic cells, and that disruption of the ATP7B-myosin Vb disrupted localization of apical membrane proteins that include interaction reduces the apical surface expression of ATP7B. P-type ATPases (Knowles et al., 2014; Müller et al., 2008). The Overexpression of the myosin Vb tail, which competes for binding apical microvilli of hepatocytes are also disrupted in MVID, and of subapical cargos to myosin Vb bound to subapical actin, disrupted clinically MVID is associated with diarrhea and cholestasis the surface expression of ATP7B, leading to reduced cellular Cu+ (Girard et al., 2014; Knowles et al., 2014; Thoeni et al., 2014). export. The myosin-Vb-dependent targeting step occurred in parallel The cholestasis may be explained as a complication secondary to with hepatocyte-like polarity. If the myosin Vb tail was expressed hyperalimentation therapy.
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus

    A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus

    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
  • 4-6 Weeks Old Female C57BL/6 Mice Obtained from Jackson Labs Were Used for Cell Isolation

    4-6 Weeks Old Female C57BL/6 Mice Obtained from Jackson Labs Were Used for Cell Isolation

    Methods Mice: 4-6 weeks old female C57BL/6 mice obtained from Jackson labs were used for cell isolation. Female Foxp3-IRES-GFP reporter mice (1), backcrossed to B6/C57 background for 10 generations, were used for the isolation of naïve CD4 and naïve CD8 cells for the RNAseq experiments. The mice were housed in pathogen-free animal facility in the La Jolla Institute for Allergy and Immunology and were used according to protocols approved by the Institutional Animal Care and use Committee. Preparation of cells: Subsets of thymocytes were isolated by cell sorting as previously described (2), after cell surface staining using CD4 (GK1.5), CD8 (53-6.7), CD3ε (145- 2C11), CD24 (M1/69) (all from Biolegend). DP cells: CD4+CD8 int/hi; CD4 SP cells: CD4CD3 hi, CD24 int/lo; CD8 SP cells: CD8 int/hi CD4 CD3 hi, CD24 int/lo (Fig S2). Peripheral subsets were isolated after pooling spleen and lymph nodes. T cells were enriched by negative isolation using Dynabeads (Dynabeads untouched mouse T cells, 11413D, Invitrogen). After surface staining for CD4 (GK1.5), CD8 (53-6.7), CD62L (MEL-14), CD25 (PC61) and CD44 (IM7), naïve CD4+CD62L hiCD25-CD44lo and naïve CD8+CD62L hiCD25-CD44lo were obtained by sorting (BD FACS Aria). Additionally, for the RNAseq experiments, CD4 and CD8 naïve cells were isolated by sorting T cells from the Foxp3- IRES-GFP mice: CD4+CD62LhiCD25–CD44lo GFP(FOXP3)– and CD8+CD62LhiCD25– CD44lo GFP(FOXP3)– (antibodies were from Biolegend). In some cases, naïve CD4 cells were cultured in vitro under Th1 or Th2 polarizing conditions (3, 4).
  • Four-Dimensional Live Imaging of Apical Biosynthetic Trafficking Reveals a Post-Golgi Sorting Role of Apical Endosomal Intermediates

    Four-Dimensional Live Imaging of Apical Biosynthetic Trafficking Reveals a Post-Golgi Sorting Role of Apical Endosomal Intermediates

    Four-dimensional live imaging of apical biosynthetic trafficking reveals a post-Golgi sorting role of apical endosomal intermediates Roland Thuenauera,b,1,2, Ya-Chu Hsua, Jose Maria Carvajal-Gonzaleza,3, Sylvie Debordea,4, Jen-Zen Chuanga, Winfried Römerc,d, Alois Sonnleitnerb, Enrique Rodriguez-Boulana,5, and Ching-Hwa Sunga,5 aMargaret M. Dyson Vision Research Institute, Weill Medical College of Cornell University, New York, NY 10065; bCenter for Advanced Bioanalysis Linz, 4020 Linz, Austria; and cInstitute of Biology II, and dBIOSS Centre for Biological Signalling Studies, Albert-Ludwigs-University Freiburg, 79104 Freiburg, Germany Edited by Keith E. Mostov, University of California School of Medicine, San Francisco, CA, and accepted by the Editorial Board January 17, 2014 (received for review March 11, 2013) Emerging data suggest that in polarized epithelial cells newly is an important regulator of biological processes that require synthesized apical and basolateral plasma membrane proteins apical trafficking, e.g., lumen formation during epithelial tubu- traffic through different endosomal compartments en route to the logenesis (11), apical secretion of discoidal/fusiform vesicles in respective cell surface. However, direct evidence for trans-endo- bladder umbrella cells (12), and apical microvillus morphogenesis somal pathways of plasma membrane proteins is still missing and and rhodopsin localization in fly photoreceptors (13). However, the mechanisms involved are poorly understood. Here, we imaged despite the physiological importance of trans-endosomal traf- the entire biosynthetic route of rhodopsin-GFP, an apical marker in ficking, the underlying mechanisms remain largely unclear. epithelial cells, synchronized through recombinant conditional ag- Previous studies on trans-endosomal trafficking in polarized gregation domains, in live Madin-Darby canine kidney cells using epithelial cells have relied on pulse chase/cell fractionation pro- spinning disk confocal microscopy.
  • Cellular and Molecular Signatures in the Disease Tissue of Early

    Cellular and Molecular Signatures in the Disease Tissue of Early

    Cellular and Molecular Signatures in the Disease Tissue of Early Rheumatoid Arthritis Stratify Clinical Response to csDMARD-Therapy and Predict Radiographic Progression Frances Humby1,* Myles Lewis1,* Nandhini Ramamoorthi2, Jason Hackney3, Michael Barnes1, Michele Bombardieri1, Francesca Setiadi2, Stephen Kelly1, Fabiola Bene1, Maria di Cicco1, Sudeh Riahi1, Vidalba Rocher-Ros1, Nora Ng1, Ilias Lazorou1, Rebecca E. Hands1, Desiree van der Heijde4, Robert Landewé5, Annette van der Helm-van Mil4, Alberto Cauli6, Iain B. McInnes7, Christopher D. Buckley8, Ernest Choy9, Peter Taylor10, Michael J. Townsend2 & Costantino Pitzalis1 1Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK. Departments of 2Biomarker Discovery OMNI, 3Bioinformatics and Computational Biology, Genentech Research and Early Development, South San Francisco, California 94080 USA 4Department of Rheumatology, Leiden University Medical Center, The Netherlands 5Department of Clinical Immunology & Rheumatology, Amsterdam Rheumatology & Immunology Center, Amsterdam, The Netherlands 6Rheumatology Unit, Department of Medical Sciences, Policlinico of the University of Cagliari, Cagliari, Italy 7Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow G12 8TA, UK 8Rheumatology Research Group, Institute of Inflammation and Ageing (IIA), University of Birmingham, Birmingham B15 2WB, UK 9Institute of
  • H-Ras Gtpase

    H-Ras Gtpase

    H-Ras GTPase Key to Understanding Cancer? Marquette University High School SMART Team: Mohammed Ayesh, Wesley Borden, Andrew Bray, Brian Digiacinto, Patrick Jordan, David Moldenhauer, Thomas Niswonger, Joseph Radke, Amit Singh, Alex Vincent, and Caleb Vogt Teachers: Keith Klestinski and David Vogt Mentor: Evgenii Kovrigin, Ph.D., Medical College of Wisconsin Abstract Cell Cycle Control The protein known as H-Ras GTPase is essential to H-ras is activated late in the G1 phase. proper biological functioning in the entire web of life. The Once H-ras is activated, the cell advances main function of this protein is giving the "stop" signal to past the G1 checkpoint and is compelled to the process of cell reproduction. Unfortunately, this protein complete mitosis. is not perfect and severe consequences, such as cancer, can arise when H-Ras GTPase malfunctions. H-Ras GTPase is a protein from the large family of enzymes that bind and split GTP. H-Ras GTPase is vital in processes like cell-to-cell communication, protein translation in ribosomes, and programmed cell death Ras GTPase Ras GDPase (apoptosis). Its main fields of operation are determining Active Inactive stem cell into specific functioning cells, as well as replicating preexisting "specialized" cells. All G domain based proteins have a universal structure and two Controlling the “Switch” between universal switch mechanisms, which consist of a mixed, six-stranded beta sheet and five alpha helices. H-Ras Active and Inactive States GTPase works by first dissociating from GDP and binding In the graphics (above and below), H-ras is shown in both © 2008, Physiomics, Corp.
  • Quantitative Live Cell Imaging Reveals Influenza Virus Manipulation Of

    Quantitative Live Cell Imaging Reveals Influenza Virus Manipulation Of

    ARTICLE https://doi.org/10.1038/s41467-019-13838-3 OPEN Quantitative live cell imaging reveals influenza virus manipulation of Rab11A transport through reduced dynein association Amar R. Bhagwat 1, Valerie Le Sage1, Eric Nturibi1, Katarzyna Kulej2, Jennifer Jones 1, Min Guo3, Eui Tae Kim 2, Benjamin A. Garcia4,5, Matthew D. Weitzman2,5,6, Hari Shroff3 & Seema S. Lakdawala 1,7* fl 1234567890():,; Assembly of infectious in uenza A viruses (IAV) is a complex process involving transport from the nucleus to the plasma membrane. Rab11A-containing recycling endosomes have been identified as a platform for intracellular transport of viral RNA (vRNA). Here, using high spatiotemporal resolution light-sheet microscopy (~1.4 volumes/second, 330 nm isotropic resolution), we quantify Rab11A and vRNA movement in live cells during IAV infection and report that IAV infection decreases speed and increases arrest of Rab11A. Unexpectedly, infection with respiratory syncytial virus alters Rab11A motion in a manner opposite to IAV, suggesting that Rab11A is a common host component that is differentially manipulated by respiratory RNA viruses. Using two-color imaging we demonstrate co-transport of Rab11A and IAV vRNA in infected cells and provide direct evidence that vRNA-associated Rab11A have altered transport. The mechanism of altered Rab11A movement is likely related to a decrease in dynein motors bound to Rab11A vesicles during IAV infection. 1 Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, 450 Technology Drive, Pittsburgh, PA 15219, USA. 2 The Children’s Hospital of Philadelphia Research Institute, 3501 Civic Center Dr., Philadelphia, PA 19104, USA. 3 Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, 13 South Drive, Building 13, Bethesda, MD 20892, USA.
  • Ncomms4301.Pdf

    Ncomms4301.Pdf

    ARTICLE Received 8 Jul 2013 | Accepted 23 Jan 2014 | Published 13 Feb 2014 DOI: 10.1038/ncomms4301 Genome-wide RNAi ionomics screen reveals new genes and regulation of human trace element metabolism Mikalai Malinouski1,2, Nesrin M. Hasan3, Yan Zhang1,4, Javier Seravalli2, Jie Lin4,5, Andrei Avanesov1, Svetlana Lutsenko3 & Vadim N. Gladyshev1 Trace elements are essential for human metabolism and dysregulation of their homoeostasis is associated with numerous disorders. Here we characterize mechanisms that regulate trace elements in human cells by designing and performing a genome-wide high-throughput siRNA/ionomics screen, and examining top hits in cellular and biochemical assays. The screen reveals high stability of the ionomes, especially the zinc ionome, and yields known regulators and novel candidates. We further uncover fundamental differences in the regulation of different trace elements. Specifically, selenium levels are controlled through the selenocysteine machinery and expression of abundant selenoproteins; copper balance is affected by lipid metabolism and requires machinery involved in protein trafficking and post-translational modifications; and the iron levels are influenced by iron import and expression of the iron/haeme-containing enzymes. Our approach can be applied to a variety of disease models and/or nutritional conditions, and the generated data set opens new directions for studies of human trace element metabolism. 1 Genetics Division, Department of Medicine, Brigham and Women’s Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA. 2 Department of Biochemistry, University of Nebraska-Lincoln, Lincoln, Nebraska 68588, USA. 3 Department of Physiology, Johns Hopkins University, Baltimore, Maryland 21205, USA. 4 Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.