Small Rab Gtpases in Intracellular Vesicle Trafficking:The Case Of
Total Page:16
File Type:pdf, Size:1020Kb
Load more
Recommended publications
-
Active Rac1 Detection Kit 2012 09/20
Active Rac1 Detection Kit 1 Kit Orders n 877-616-CELL (2355) (30 immunoprecipitations) [email protected] Support n 877-678-TECH (8324) [email protected] Web n www.cellsignal.com rev. 09/24/20 #8815 For Research Use Only. Not For Use In Diagnostic Procedures. Species Cross-Reactivity: H, M Components Ship As: 11894S Item # Kit Quantity Storage Temp Description: The Active Rac1 Detection Kit provides all GTPgS 11521 1 X 50 µl –80°C reagents necessary for measuring activation of Rac1 GTPase in the cell. GST-PAK1-PBD fusion protein is used to bind GDP 11522 1 X 50 µl –80°C the activated form of GTP-bound Rac1, which can then be Components Ship As: 11894S Item # Kit Quantity Storage Temp immunoprecipitated with glutathione resin. Rac1 activation levels are then determined by western blot using a Rac1 GST-Human PAK1-PBD 8659 1 X 600 µg –20°C Mouse mAb. Rac1 Mouse mAb 8631 1 X 50 µl –20°C Specificity/Sensitivity: Active Rac1 Detection Kit detects Components Ship As: 11860S Item # Kit Quantity Storage Temp endogenous levels of GTP-bound (active) Rac1 as shown in Lysis/Binding/Wash Buffer 11524 1 X 100 mL 4°C Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect Glutathione Resin 11523 1 X 3 ml 4°C homologous proteins from other species. SDS Sample Buffer 11525 1 X 1.5 ml 4°C Background: The Ras superfamily of small GTP-binding Spin Cup and Collection Tubes 11526 1 X 30 vial RT proteins (G proteins) comprise a large class of proteins (over 150 members) that can be classified into at least five families based on their sequence and functional similarities: 1 2 3 4 Figure 1. -
Rac1 Accumulates in the Nucleus During the G2 Phase of the Cell
Published Online: 28 April, 2008 | Supp Info: http://doi.org/10.1083/jcb.200801047 JCB: ARTICLE Downloaded from jcb.rupress.org on May 9, 2018 Rac1 accumulates in the nucleus during the G2 phase of the cell cycle and promotes cell division David Michaelson , 2,5 Wasif Abidi , 2,5 Daniele Guardavaccaro , 4,5 Mo Zhou, 3,5 Ian Ahearn , 3,5 Michele Pagano , 4,5 and Mark R. Philips 1,2,3,5 1 Department of Medicine, 2 Department of Cell Biology, 3 Department of Pharmacology, 4 Department of Pathology, and the 5 New York University Cancer Institute, New York University School of Medicine, New York, NY 10016 ac1 regulates a wide variety of cellular processes. zation of cells stably expressing low levels of GFP-Rac1, The polybasic region of the Rac1 C terminus func- and time-lapse microscopy of asynchronous cells revealed Rtions both as a plasma membrane – targeting motif Rac1 accumulation in the nucleus in late G2 and exclu- and a nuclear localization sequence (NLS). We show that sion in early G1. Although constitutively active Rac1 re- a triproline N-terminal to the polybasic region contributes stricted to the cytoplasm inhibited cell division, activated to the NLS, which is cryptic in the sense that it is strongly Rac1 expressed constitutively in the nucleus increased the inhibited by geranylgeranylation of the adjacent cysteine. mitotic rate. These results show that Rac1 cycles in and out Subcellular fractionation demonstrated endogenous Rac1 of the nucleus during the cell cycle and thereby plays a in the nucleus and Triton X-114 partition revealed that this role in promoting cell division. -
Rab3a As a Modulator of Homeostatic Synaptic Plasticity
Wright State University CORE Scholar Browse all Theses and Dissertations Theses and Dissertations 2014 Rab3A as a Modulator of Homeostatic Synaptic Plasticity Andrew G. Koesters Wright State University Follow this and additional works at: https://corescholar.libraries.wright.edu/etd_all Part of the Biomedical Engineering and Bioengineering Commons Repository Citation Koesters, Andrew G., "Rab3A as a Modulator of Homeostatic Synaptic Plasticity" (2014). Browse all Theses and Dissertations. 1246. https://corescholar.libraries.wright.edu/etd_all/1246 This Dissertation is brought to you for free and open access by the Theses and Dissertations at CORE Scholar. It has been accepted for inclusion in Browse all Theses and Dissertations by an authorized administrator of CORE Scholar. For more information, please contact [email protected]. RAB3A AS A MODULATOR OF HOMEOSTATIC SYNAPTIC PLASTICITY A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy By ANDREW G. KOESTERS B.A., Miami University, 2004 2014 Wright State University WRIGHT STATE UNIVERSITY GRADUATE SCHOOL August 22, 2014 I HEREBY RECOMMEND THAT THE DISSERTATION PREPARED UNDER MY SUPERVISION BY Andrew G. Koesters ENTITLED Rab3A as a Modulator of Homeostatic Synaptic Plasticity BE ACCEPTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF Doctor of Philosophy. Kathrin Engisch, Ph.D. Dissertation Director Mill W. Miller, Ph.D. Director, Biomedical Sciences Ph.D. Program Committee on Final Examination Robert E. W. Fyffe, Ph.D. Vice President for Research Dean of the Graduate School Mark Rich, M.D./Ph.D. David Ladle, Ph.D. F. Javier Alvarez-Leefmans, M.D./Ph.D. Lynn Hartzler, Ph.D. -
Trafficking Routes to the Plant Vacuole: Connecting Alternative and Classical Pathways
This is a repository copy of Trafficking routes to the plant vacuole: connecting alternative and classical pathways. White Rose Research Online URL for this paper: http://eprints.whiterose.ac.uk/124374/ Version: Accepted Version Article: Di Sansebastiano, GP, Barozzi, F, Piro, G et al. (2 more authors) (2018) Trafficking routes to the plant vacuole: connecting alternative and classical pathways. Journal of Experimental Botany, 69 (1). pp. 79-90. ISSN 0022-0957 https://doi.org/10.1093/jxb/erx376 © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology. This is an author produced version of a paper published in Journal of Experimental Botany. Uploaded in accordance with the publisher's self-archiving policy. Reuse Items deposited in White Rose Research Online are protected by copyright, with all rights reserved unless indicated otherwise. They may be downloaded and/or printed for private study, or other acts as permitted by national copyright laws. The publisher or other rights holders may allow further reproduction and re-use of the full text version. This is indicated by the licence information on the White Rose Research Online record for the item. Takedown If you consider content in White Rose Research Online to be in breach of UK law, please notify us by emailing [email protected] including the URL of the record and the reason for the withdrawal request. [email protected] https://eprints.whiterose.ac.uk/ 1 Trafficking routes to the Plant Vacuole: 2 connecting alternative and classical pathways. 3 4 Gian Pietro Di Sansebastiano1*, Fabrizio Barozzi1, Gabriella Piro1, Jurgen Denecke2 and 5 Carine de Marcos Lousa2,3 * 6 1.! DiSTeBA (Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali), 7 University of Salento, Campus ECOTEKNE, 73100 Lecce, Italy; 8 [email protected] and [email protected]. -
Human Placenta Exosomes: Biogenesis, Isolation, Composition, and Prospects for Use in Diagnostics
International Journal of Molecular Sciences Review Human Placenta Exosomes: Biogenesis, Isolation, Composition, and Prospects for Use in Diagnostics Evgeniya E. Burkova 1,* , Sergey E. Sedykh 1,2 and Georgy A. Nevinsky 1,2 1 SB RAS Institute of Chemical Biology and Fundamental Medicine, 630090 Novosibirsk, Russia; [email protected] (S.E.S.); [email protected] (G.A.N.) 2 Department of Natural Sciences, Novosibirsk State University, 630090 Novosibirsk, Russia * Correspondence: [email protected]; Tel.: +7-(383)-363-51-27 Abstract: Exosomes are 40–100 nm nanovesicles participating in intercellular communication and transferring various bioactive proteins, mRNAs, miRNAs, and lipids. During pregnancy, the placenta releases exosomes into the maternal circulation. Placental exosomes are detected in the maternal blood even in the first trimester of pregnancy and their numbers increase significantly by the end of pregnancy. Exosomes are necessary for the normal functioning of the placenta and fetal development. Effects of exosomes on target cells depend not only on their concentration but also on their intrinsic components. The biochemical composition of the placental exosomes may cause various complications of pregnancy. Some studies relate the changes in the composition of nanovesicles to placental dysfunction. Isolation of placental exosomes from the blood of pregnant women and the study of protein, lipid, and nucleic composition can lead to the development of methods for early diagnosis of pregnancy pathologies. This review describes the biogenesis of exosomes, methods Citation: Burkova, E.E.; Sedykh, S.E.; of their isolation, analyzes their biochemical composition, and considers the prospects for using Nevinsky, G.A. Human Placenta exosomes to diagnose pregnancy pathologies. -
Sorting Nexins in Protein Homeostasis Sara E. Hanley1,And Katrina F
Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 6 November 2020 doi:10.20944/preprints202011.0241.v1 Sorting nexins in protein homeostasis Sara E. Hanley1,and Katrina F. Cooper2* 1Department of Molecular Biology, Graduate School of Biomedical Sciences, Rowan University, Stratford, NJ, 08084, USA 1 [email protected] 2 [email protected] * [email protected] Tel: +1 (856)-566-2887 1Department of Molecular Biology, Graduate School of Biomedical Sciences, Rowan University, Stratford, NJ, 08084, USA Abstract: Sorting nexins (SNXs) are a highly conserved membrane-associated protein family that plays a role in regulating protein homeostasis. This family of proteins is unified by their characteristic phox (PX) phosphoinositides binding domain. Along with binding to membranes, this family of SNXs also comprises a diverse array of protein-protein interaction motifs that are required for cellular sorting and protein trafficking. SNXs play a role in maintaining the integrity of the proteome which is essential for regulating multiple fundamental processes such as cell cycle progression, transcription, metabolism, and stress response. To tightly regulate these processes proteins must be expressed and degraded in the correct location and at the correct time. The cell employs several proteolysis mechanisms to ensure that proteins are selectively degraded at the appropriate spatiotemporal conditions. SNXs play a role in ubiquitin-mediated protein homeostasis at multiple levels including cargo localization, recycling, degradation, and function. In this review, we will discuss the role of SNXs in three different protein homeostasis systems: endocytosis lysosomal, the ubiquitin-proteasomal, and the autophagy-lysosomal system. The highly conserved nature of this protein family by beginning with the early research on SNXs and protein trafficking in yeast and lead into their important roles in mammalian systems. -
Direct Lysosome-Based Autophagy of Lipid Droplets in Hepatocytes
Direct lysosome-based autophagy of lipid droplets in hepatocytes Ryan J. Schulzea,b,1, Eugene W. Kruegera,b,1, Shaun G. Wellera,b, Katherine M. Johnsona,b, Carol A. Caseyc, Micah B. Schotta,b, and Mark A. McNivena,b,2 aDepartment of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905; bDivision of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN 55905; and cDepartment of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68198 Edited by Tobias C. Walther, Harvard School of Public Health, Boston, MA, and accepted by Editorial Board Member Joseph L. Goldstein October 13, 2020 (received for review June 4, 2020) Hepatocytes metabolize energy-rich cytoplasmic lipid droplets (LDs) process that appears to play an especially important role in the in the lysosome-directed process of autophagy. An organelle- degradation of hepatocellular LDs (15). A selective form of LD- selective form of this process (macrolipophagy) results in the engulf- centric autophagy known as lipophagy is thought to involve the ment of LDs within double-membrane delimited structures (auto- recognition of as-of-yet unidentified LD-specific receptors to phagosomes) before lysosomal fusion. Whether this is an promote the localized assembly and extension of a sequestering exclusive autophagic mechanism used by hepatocytes to catabolize phagophore around the perimeter of the LD (16, 17). How this LDs is unclear. It is also unknown whether lysosomes alone might be phagophore is targeted to (and extended around) the LD surface to sufficient to mediate LD turnover in the absence of an autophago- facilitate lipophagy remains unclear. Once fully enclosed, the somal intermediate. -
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. -
Glioma Cell Secretion: a Driver of Tumor Progression and a Potential Therapeutic Target Damian A
Published OnlineFirst October 17, 2018; DOI: 10.1158/0008-5472.CAN-18-0345 Cancer Review Research Glioma Cell Secretion: A Driver of Tumor Progression and a Potential Therapeutic Target Damian A. Almiron Bonnin1,2, Matthew C. Havrda1,2, and Mark A. Israel1,2,3 Abstract Cellular secretion is an important mediator of cancer progres- ple oncogenic pathologies. In this review, we describe tumor cell sion. Secreted molecules in glioma are key components of secretion in high-grade glioma and highlight potential novel complex autocrine and paracrine pathways that mediate multi- therapeutic opportunities. Cancer Res; 78(21); 6031–9. Ó2018 AACR. Introduction Glioma-Secreted Molecules Impact Disease Glial cells in the central nervous system (CNS) provide trophic Progression support for neurons (1). In glial tumors, this trophic support is Glioma cells modify their microenvironment by introducing dysregulated creating a pro-oncogenic microenvironment medi- diverse molecules into the extracellular space (Table 1). To exem- ated by a heterogeneous array of molecules secreted into the plify the pro-oncogenic role that secreted molecules can have on – extracellular space (2 15). The glioma secretome includes pro- glioma pathology, we review the functional impact of specific teins, nucleic acids, and metabolites that are often overexpressed cytokines, metabolites, and nucleic acids on glioma biology. By in malignant tissue and contribute to virtually every aspect of describing some of the potent antitumorigenic effects observed in – cancer pathology (Table 1; Fig. 1; refs. 2 15), providing a strong preclinical therapeutic studies targeting tumor cell secretion, we – rationale to target the cancer cell secretory mechanisms. also highlight how blocking secreted molecules might be of fi Although the speci c mechanisms regulating secretion in therapeutic impact in gliomas, as well as other tumors. -
Ubiquitylome Profiling of Parkin-Null Brain Reveals Dysregulation Of
Neurobiology of Disease 127 (2019) 114–130 Contents lists available at ScienceDirect Neurobiology of Disease journal homepage: www.elsevier.com/locate/ynbdi Ubiquitylome profiling of Parkin-null brain reveals dysregulation of calcium T homeostasis factors ATP1A2, Hippocalcin and GNA11, reflected by altered firing of noradrenergic neurons Key J.a,1, Mueller A.K.b,1, Gispert S.a, Matschke L.b, Wittig I.c, Corti O.d,e,f,g, Münch C.h, ⁎ ⁎ Decher N.b, , Auburger G.a, a Exp. Neurology, Goethe University Medical School, 60590 Frankfurt am Main, Germany b Institute for Physiology and Pathophysiology, Vegetative Physiology and Marburg Center for Mind, Brain and Behavior - MCMBB; Clinic for Neurology, Philipps-University Marburg, 35037 Marburg, Germany c Functional Proteomics, SFB 815 Core Unit, Goethe University Medical School, 60590 Frankfurt am Main, Germany d Institut du Cerveau et de la Moelle épinière, ICM, Paris, F-75013, France e Inserm, U1127, Paris, F-75013, France f CNRS, UMR 7225, Paris, F-75013, France g Sorbonne Universités, Paris, F-75013, France h Institute of Biochemistry II, Goethe University Medical School, 60590 Frankfurt am Main, Germany ARTICLE INFO ABSTRACT Keywords: Parkinson's disease (PD) is the second most frequent neurodegenerative disorder in the old population. Among Parkinson's disease its monogenic variants, a frequent cause is a mutation in the Parkin gene (Prkn). Deficient function of Parkin Mitochondria triggers ubiquitous mitochondrial dysfunction and inflammation in the brain, but it remains unclear howse- Parkin lective neural circuits become vulnerable and finally undergo atrophy. Ubiquitin We attempted to go beyond previous work, mostly done in peripheral tumor cells, which identified protein Calcium targets of Parkin activity, an ubiquitin E3 ligase. -
Supporting Information
Supporting Information Edgar et al. 10.1073/pnas.1601895113 SI Methods (Actimetrics), and recordings were analyzed using LumiCycle Mice. Sample size was determined using the resource equation: Data Analysis software (Actimetrics). E (degrees of freedom in ANOVA) = (total number of exper- – Cell Cycle Analysis of Confluent Cell Monolayers. NIH 3T3, primary imental animals) (number of experimental groups), with −/− sample size adhering to the condition 10 < E < 20. For com- WT, and Bmal1 fibroblasts were sequentially transduced − − parison of MuHV-4 and HSV-1 infection in WT vs. Bmal1 / with lentiviral fluorescent ubiquitin-based cell cycle indicators mice at ZT7 (Fig. 2), the investigator did not know the genotype (FUCCI) mCherry::Cdt1 and amCyan::Geminin reporters (32). of the animals when conducting infections, bioluminescence Dual reporter-positive cells were selected by FACS (Influx Cell imaging, and quantification. For bioluminescence imaging, Sorter; BD Biosciences) and seeded onto 35-mm dishes for mice were injected intraperitoneally with endotoxin-free lucif- subsequent analysis. To confirm that expression of mCherry:: Cdt1 and amCyan::Geminin correspond to G1 (2n DNA con- erin (Promega E6552) using 2 mg total per mouse. Following < ≤ anesthesia with isofluorane, they were scanned with an IVIS tent) and S/G2 (2 n 4 DNA content) cell cycle phases, Lumina (Caliper Life Sciences), 15 min after luciferin admin- respectively, cells were stained with DNA dye DRAQ5 (abcam) and analyzed by flow cytometry (LSR-Fortessa; BD Biosci- istration. Signal intensity was quantified using Living Image ences). To examine dynamics of replicative activity under ex- software (Caliper Life Sciences), obtaining maximum radiance perimental confluent conditions, synchronized FUCCI reporter for designated regions of interest (photons per second per − − − monolayers were observed by time-lapse live cell imaging over square centimeter per Steradian: photons·s 1·cm 2·sr 1), relative 3 d (Nikon Eclipse Ti-E inverted epifluorescent microscope). -
ADP-Ribosylation Factor, a Small GTP-Binding Protein, Is Required for Binding of the Coatomer Protein Fl-COP to Golgi Membranes JULIE G
Proc. Natl. Acad. Sci. USA Vol. 89, pp. 6408-6412, July 1992 Biochemistry ADP-ribosylation factor, a small GTP-binding protein, is required for binding of the coatomer protein fl-COP to Golgi membranes JULIE G. DONALDSON*, DAN CASSEL*t, RICHARD A. KAHN*, AND RICHARD D. KLAUSNER* *Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, and tLaboratory of Biological Chemistry, Division of Cancer Treatment, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 Communicated by Marc Kirschner, April 20, 1992 (receivedfor review February 11, 1992) ABSTRACT The coatomer is a cytosolic protein complex localized to the Golgi complex, although their functions have that reversibly associates with Golgi membranes and is Impli- not been defined. Distinct among these proteins is the ADP- cated in modulating Golgi membrane transport. The associa- ribosylation factor (ARF), originally identified as a cofactor tion of 13-COP, a component of coatomer, with Golgi mem- required for in vitro cholera toxin-catalyzed ADP- branes is enhanced by guanosine 5'-[v-thioltriphosphate ribosylation of the a subunit of the trimeric GTP-binding (GTP[yS]), a nonhydrolyzable analogue of GTP, and by a protein G, (G,.) (19). ARF is an abundant cytosolic protein mixture of aluminum and fluoride ions (Al/F). Here we show that reversibly associates with Golgi membranes (20, 21). that the ADP-ribosylation factor (ARF) is required for the ARF has been shown to be present on Golgi coated vesicles binding of (-COP. Thus, 13-COP contained in a coatomer generated in the presence of GTP[yS], but it is not a com- fraction that has been resolved from ARF does not bind to Golgi ponent of the cytosolic coatomer (22).