This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Al-Awar, O.
Right arrow Articles by Donaldson, J. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Al-Awar, O.
Right arrow Articles by Donaldson, J. G.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, August 2000, p. 5998-6007, Vol. 20, No. 16
0270-7306/00/$04.00+0

Separation of Membrane Trafficking and Actin Remodeling Functions of ARF6 with an Effector Domain Mutant

Omayma Al-Awar, Harish Radhakrishna,dagger Natasha N. Powell, and Julie G. Donaldson*

Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892

Received 21 December 1999/Returned for modification 9 February 2000/Accepted 15 May 2000

The ADP-ribosylation factor 6 (ARF6) GTPase has a dual function in cells, regulating membrane traffic and organizing cortical actin. ARF6 activation is required for recycling of the endosomal membrane back to the plasma membrane (PM) and also for ruffling at the PM induced by Rac. Additionally, ARF6 at the PM induces the formation of actin-containing protrusions. To identify sequences in ARF6 that are necessary for these distinct functions, we examined the behavior of a chimeric protein of ARF1 and ARF6. The 1-6 chimera (with the amino half of ARF1 and the carboxyl half of ARF6) localized like ARF6 in HeLa cells and moved between the endosome and PM, but it did not form protrusions, an ARF6 effector function. Two residues in the amino-terminal half of ARF6, Q37 and S38, when substituted into the 1-6 chimera allowed protrusion formation, whereas removal of these residues from ARF6 resulted in an inability to form protrusions. Interestingly, expression of 1-6 in cells selectively inhibited protrusions induced by wild-type ARF6 but had no effect on ARF6-regulated membrane movement or Rac-induced ruffling. Thus, we have uncoupled two functions of ARF6, one involved in membrane trafficking, which is necessary for Rac ruffling, and another involved in protrusion formation.

* Corresponding author. Mailing address: Center Dr. MSC 0301, Bldg. 3, Room B1-22, Bethesda, MD 20892. Phone: (301) 402-2907. Fax: (301) 402-1519. E-mail: jdonalds{at}helix.nih.gov.

dagger Present address: School of Biology, Georgia Institute of Technology, Atlanta, GA 30332-0363.

Molecular and Cellular Biology, August 2000, p. 5998-6007, Vol. 20, No. 16
0270-7306/00/$04.00+0


This article has been cited by other articles:

  • Klein, S., Partisani, M., Franco, M., Luton, F. (2008). EFA6 Facilitates the Assembly of the Tight Junction by Coordinating an Arf6-dependent and -independent Pathway. J. Biol. Chem. 283: 30129-30138 [Abstract] [Full Text]  
  • Price, H. P., Stark, M., Smith, D. F. (2007). Trypanosoma brucei ARF1 Plays a Central Role in Endocytosis and Golgi-Lysosome Trafficking. Mol. Biol. Cell 18: 864-873 [Abstract] [Full Text]  
  • Goldfinger, L. E., Ptak, C., Jeffery, E. D., Shabanowitz, J., Hunt, D. F., Ginsberg, M. H. (2006). RLIP76 (RalBP1) is an R-Ras effector that mediates adhesion-dependent Rac activation and cell migration. JCB 174: 877-888 [Abstract] [Full Text]  
  • Klein, S., Franco, M., Chardin, P., Luton, F. (2006). Role of the Arf6 GDP/GTP Cycle and Arf6 GTPase-activating Proteins in Actin Remodeling and Intracellular Transport. J. Biol. Chem. 281: 12352-12361 [Abstract] [Full Text]  
  • Li, M., Ng, S. S.-m., Wang, J., Lai, L., Yi Leung, S., Franco, M., Peng, Y., He, M.-l., Kung, H.-f., Lin, M. C.-m. (2006). EFA6A Enhances Glioma Cell Invasion through ADP Ribosylation Factor 6/Extracellular Signal-Regulated Kinase Signaling. Cancer Res. 66: 1583-1590 [Abstract] [Full Text]  
  • Robertson, S. E., Setty, S. R. G., Sitaram, A., Marks, M. S., Lewis, R. E., Chou, M. M. (2006). Extracellular Signal-regulated Kinase Regulates Clathrin-independent Endosomal Trafficking. Mol. Biol. Cell 17: 645-657 [Abstract] [Full Text]  
  • Balana, M. E., Niedergang, F., Subtil, A., Alcover, A., Chavrier, P., Dautry-Varsat, A. (2005). ARF6 GTPase controls bacterial invasion by actin remodelling. J. Cell Sci. 118: 2201-2210 [Abstract] [Full Text]  
  • Tanabe, K., Torii, T., Natsume, W., Braesch-Andersen, S., Watanabe, T., Satake, M. (2005). A Novel GTPase-activating Protein for ARF6 Directly Interacts with Clathrin and Regulates Clathrin-dependent Endocytosis. Mol. Biol. Cell 16: 1617-1628 [Abstract] [Full Text]  
  • Honda, A., Al-Awar, O. S., Hay, J. C., Donaldson, J. G. (2005). Targeting of Arf-1 to the early Golgi by membrin, an ER-Golgi SNARE. JCB 168: 1039-1051 [Abstract] [Full Text]  
  • LI, Y., KELLY, W. G., LOGSDON, J. M. JR, SCHURKO, A. M., HARFE, B. D., HILL-HARFE, K. L., KAHN, R. A. (2004). Functional genomic analysis of the ADP-ribosylation factor family of GTPases: phylogeny among diverse eukaryotes and function in C. elegans. FASEB J. 18: 1834-1850 [Abstract] [Full Text]  
  • Hashimoto, S., Hashimoto, A., Yamada, A., Kojima, C., Yamamoto, H., Tsutsumi, T., Higashi, M., Mizoguchi, A., Yagi, R., Sabe, H. (2004). A Novel Mode of Action of an ArfGAP, AMAP2/PAG3/Pap{alpha}, in Arf6 Function. J. Biol. Chem. 279: 37677-37684 [Abstract] [Full Text]  
  • Prigent, M., Dubois, T., Raposo, G., Derrien, V., Tenza, D., Rosse, C., Camonis, J., Chavrier, P. (2003). ARF6 controls post-endocytic recycling through its downstream exocyst complex effector. JCB 163: 1111-1121 [Abstract] [Full Text]  
  • Donaldson, J. G. (2003). Multiple Roles for Arf6: Sorting, Structuring, and Signaling at the Plasma Membrane. J. Biol. Chem. 278: 41573-41576 [Full Text]  
  • Sabe, H. (2003). Requirement for Arf6 in Cell Adhesion, Migration, and Cancer Cell Invasion. J Biochem 134: 485-489 [Abstract] [Full Text]  
  • Kim, S., Ko, J., Shin, H., Lee, J.-R., Lim, C., Han, J.-H., Altrock, W. D., Garner, C. C., Gundelfinger, E. D., Premont, R. T., Kaang, B.-K., Kim, E. (2003). The GIT Family of Proteins Forms Multimers and Associates with the Presynaptic Cytomatrix Protein Piccolo. J. Biol. Chem. 278: 6291-6300 [Abstract] [Full Text]  
  • Lee, M. H., Min, M. K., Lee, Y. J., Jin, J. B., Shin, D. H., Kim, D. H., Lee, K.-H., Hwang, I. (2002). ADP-Ribosylation Factor 1 of Arabidopsis Plays a Critical Role in Intracellular Trafficking and Maintenance of Endoplasmic Reticulum Morphology in Arabidopsis. Plant Physiol. 129: 1507-1520 [Abstract] [Full Text]  
  • Schweitzer, J. K., D'Souza-Schorey, C. (2002). Localization and Activation of the ARF6 GTPase during Cleavage Furrow Ingression and Cytokinesis. J. Biol. Chem. 277: 27210-27216 [Abstract] [Full Text]  
  • Brown, M. C., West, K. A., Turner, C. E. (2002). Paxillin-dependent Paxillin Kinase Linker and p21-Activated Kinase Localization to Focal Adhesions Involves a Multistep Activation Pathway. Mol. Biol. Cell 13: 1550-1565 [Abstract] [Full Text]  
  • Lu, L., Horstmann, H., Ng, C., Hong, W. (2002). Regulation of Golgi structure and function by ARF-like protein 1 (Arl1). J. Cell Sci. 114: 4543-4555 [Abstract] [Full Text]  
  • Shinotsuka, C., Yoshida, Y., Kawamoto, K., Takatsu, H., Nakayama, K. (2002). Overexpression of an ADP-ribosylation Factor-Guanine Nucleotide Exchange Factor, BIG2, Uncouples Brefeldin A-induced Adaptor Protein-1 Coat Dissociation and Membrane Tubulation. J. Biol. Chem. 277: 9468-9473 [Abstract] [Full Text]  
  • Laporte, J., Blondeau, F., Gansmuller, A., Lutz, Y., Vonesch, J.-L., Mandel, J.-L. (2002). The PtdIns3P phosphatase myotubularin is a cytoplasmic protein that also localizes to Rac1-inducible plasma membrane ruffles. J. Cell Sci. 115: 3105-3117 [Abstract] [Full Text]  
  • Bose, A., Cherniack, A. D., Langille, S. E., Nicoloro, S. M. C., Buxton, J. M., Park, J. G., Chawla, A., Czech, M. P. (2001). G{alpha}11 Signaling through ARF6 Regulates F-Actin Mobilization and GLUT4 Glucose Transporter Translocation to the Plasma Membrane. Mol. Cell. Biol. 21: 5262-5275 [Abstract] [Full Text]  
  • Lawrence, J. T. R., Birnbaum, M. J. (2001). ADP-Ribosylation Factor 6 Delineates Separate Pathways Used by Endothelin 1 and Insulin for Stimulating Glucose Uptake in 3T3-L1 Adipocytes. Mol. Cell. Biol. 21: 5276-5285 [Abstract] [Full Text]  
  • West, K. A., Zhang, H., Brown, M. C., Nikolopoulos, S. N., Riedy, M.C., Horwitz, A. F., Turner, C. E. (2001). The LD4 motif of paxillin regulates cell spreading and motility through an interaction with paxillin kinase linker (PKL). JCB 154: 161-176 [Abstract] [Full Text]  
  • Uchida, H., Kondo, A., Yoshimura, Y., Mazaki, Y., Sabe, H. (2001). Pag3/Pap{alpha}/Kiaa0400, a Gtpase-Activating Protein for Adp-Ribosylation Factor (Arf), Regulates Arf6 in Fc{gamma} Receptor-Mediated Phagocytosis of Macrophages. JEM 193: 955-966 [Abstract] [Full Text]  
  • Yahara, N., Ueda, T., Sato, K., Nakano, A. (2001). Multiple Roles of Arf1 GTPase in the Yeast Exocytic and Endocytic Pathways. Mol. Biol. Cell 12: 221-238 [Abstract] [Full Text]  
  • Jobling, M. G., Holmes, R. K. (2000). Identification of motifs in cholera toxin A1 polypeptide that are required for its interaction with human ADP-ribosylation factor 6 in a bacterial two-hybrid system. Proc. Natl. Acad. Sci. USA 10.1073/pnas.011442598v1 [Abstract] [Full Text]  
  • Vitale, N., Ferrans, V. J., Moss, J., Vaughan, M. (2000). Identification of Lysosomal and Golgi Localization Signals in GAP and ARF Domains of ARF Domain Protein 1. Mol. Cell. Biol. 20: 7342-7352 [Abstract] [Full Text]  
  • Jobling, M. G., Holmes, R. K. (2000). Identification of motifs in cholera toxin A1 polypeptide that are required for its interaction with human ADP-ribosylation factor 6 in a bacterial two-hybrid system. Proc. Natl. Acad. Sci. USA 97: 14662-14667 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»