HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 278, Issue 44, 43460-43469, October 31, 2003

This Article Full Text Full Text (PDF) All Versions of this Article: 278/44/43460    most recent M304078200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Venkateswarlu, K. Search for Related Content PubMed PubMed Citation Articles by Venkateswarlu, K. Social Bookmarking                      What's this?

Interaction Protein for Cytohesin Exchange Factors 1 (IPCEF1) Binds Cytohesin 2 and Modifies Its Activity^*

Kanamarlapudi Venkateswarlu

From the Department of Pharmacology, The University of Bristol, Bristol BS8 1TD, United Kingdom

The ADP-ribosylation factor 6 (ARF6) small GTPase functions as a GDP/GTP-regulated switch in the pathways that stimulate actin reorganization and membrane ruffling. The formation of active ARF6_GTP is stimulated by guanine nucleotide exchange factors (GEFs) such as cytohesins, which translocate to the plasma membrane in agonist-stimulated cells by binding the lipid second messenger phosphatidylinositol 3,4,5-trisphosphate through the pleckstrin homology domain with subsequent ARF6 activation. Using cytohesin 2 as bait in yeast two-hybrid screening, we have isolated a cDNA encoding a protein termed interaction protein for cytohesin exchange factors 1 (IPCEF1). Using yeast two-hybrid and glutathione S-transferase pull-down assays coupled with deletion mutational analysis, the specific domains required for the cytohesin 2-IPCEF1 interaction were mapped to the coiled-coil domain of cytohesin 2 and the C-terminal 121 amino acids of IPCEF1. IPCEF1 also interacts with the other members of the cytohesin family of ARF GEFs, suggesting that the interaction with IPCEF1 is highly conserved among the cytohesin family of ARF GEFs. The interaction of cytohesin 2 and IPCEF1 in mammalian cells was demonstrated by immunoprecipitation. Immunofluorescence analysis revealed that IPCEF1 co-localizes with cytohesin 2 to the cytosol in unstimulated cells and translocates to the plasma membrane via binding to cytohesin 2 in epidermal growth factor-stimulated cells. However, a deletion mutant of IPCEF1 that lacks the cytohesin 2 binding site failed to co-migrate with cytohesin 2 to the membrane in stimulated cells. The functional significance of the IPCEF1-cytohesin 2 interaction is demonstrated by showing that IPCEF1 increases the in vitro and in vivo stimulation of ARF_GTP formation by cytohesin 2.

Received for publication, April 17, 2003 , and in revised form, August 14, 2003.

The nucleotide sequence(s) reported in this paper has been submitted to the DDBJ/GenBank^TM/EBI Data Bank with accession number(s) AJ536192.

^* This work was funded by the Biotechnology and Biological Science Research Council (BBSRC) and the Royal Society UK. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Recipient of a David Phillips Research Fellowship from the BBSRC. To whom correspondence should be addressed: Dept. of Pharmacology, School of Medical Sciences, The University of Bristol, University Walk, Bristol BS8 1TD, UK. Tel.: 44-117-9289702; Fax: 44-117-9250168; E-mail: k.venkateswarlu{at}bristol.ac.uk.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				C.-C. Li, T.-C. Chiang, T.-S. Wu, G. Pacheco-Rodriguez, J. Moss, and F.-J. S. Lee ARL4D Recruits Cytohesin-2/ARNO to Modulate Actin Remodeling Mol. Biol. Cell, November 1, 2007; 18(11): 4420 - 4437. [Abstract] [Full Text] [PDF]

				K. Venkateswarlu, K. G. Brandom, and H. Yun PI-3-kinase-dependent membrane recruitment of centaurin-{alpha}2 is essential for its effect on ARF6-mediated actin cytoskeleton reorganisation J. Cell Sci., March 1, 2007; 120(5): 792 - 801. [Abstract] [Full Text] [PDF]

				M. Shmuel, L. C. Santy, S. Frank, D. Avrahami, J. E. Casanova, and Y. Altschuler ARNO through Its Coiled-coil Domain Regulates Endocytosis at the Apical Surface of Polarized Epithelial Cells J. Biol. Chem., May 12, 2006; 281(19): 13300 - 13308. [Abstract] [Full Text] [PDF]

				K. Venkateswarlu, T. Hanada, and A. H. Chishti Centaurin-{alpha}1 interacts directly with kinesin motor protein KIF13B J. Cell Sci., June 1, 2005; 118(11): 2471 - 2484. [Abstract] [Full Text] [PDF]

				J. Lawrence, S. J. Mundell, H. Yun, E. Kelly, and K. Venkateswarlu Centaurin-{alpha}1, an ADP-Ribosylation Factor 6 GTPase Activating Protein, Inhibits {beta}2-Adrenoceptor Internalization Mol. Pharmacol., June 1, 2005; 67(6): 1822 - 1828. [Abstract] [Full Text] [PDF]

				R. Cox, R. J Mason-Gamer, C. L. Jackson, and N. Segev Phylogenetic Analysis of Sec7-Domain-containing Arf Nucleotide Exchangers Mol. Biol. Cell, April 1, 2004; 15(4): 1487 - 1505. [Abstract] [Full Text] [PDF]

				K. Venkateswarlu, K. G. Brandom, and J. L. Lawrence Centaurin-{alpha}1 Is an in Vivo Phosphatidylinositol 3,4,5-Trisphosphate-dependent GTPase-activating Protein for ARF6 That Is Involved in Actin Cytoskeleton Organization J. Biol. Chem., February 20, 2004; 279(8): 6205 - 6208. [Abstract] [Full Text] [PDF]