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

J. Biol. Chem., Vol. 278, Issue 40, 38786-38795, October 3, 2003

This Article Full Text Full Text (PDF) All Versions of this Article: 278/40/38786    most recent M306864200v1 Alert me when this article is cited 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 Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Whitley, P. Articles by Holman, G. D. Search for Related Content PubMed PubMed Citation Articles by Whitley, P. Articles by Holman, G. D. Social Bookmarking                      What's this?

Identification of Mammalian Vps24p as an Effector of Phosphatidylinositol 3,5-Bisphosphate-dependent Endosome Compartmentalization^*

Paul Whitley  , Barbara J. Reaves , Makoto Hashimoto , Andrew M. Riley ¶, Barry V. L. Potter ¶ and Geoffrey D. Holman 

From the Department of Biology and Biochemistry and the ^¶Wolfson Laboratory of Medicinal Chemistry, Department of Pharmacy and Pharmacology, University of Bath, BA2 7AY, United Kingdom

Phosphatidylinositol 3,5-bisphosphate is a membrane lipid found in all eukaryotes so far studied but downstream effector proteins of this lipid have yet to be identified. Here we report the use of cDNA phage libraries in conjunction with synthetic biotinylated derivatives of phosphatidylinositol 3,5-bisphosphate in the identification of a mammalian phosphatidylinositol 3,5-bisphosphate-binding protein, mVps24p. This protein is orthologous to the Saccharomyces cerevisiae protein, Vps24p, a class-E vacuolar protein-sorting protein. Using in vitro liposome binding and competition assays, we demonstrate that mVps24p selectively binds to phosphatidylinositol 3,5-bisphosphate and phosphatidylinositol 3,4-bisphosphate in preference to other phosphoinositides tested. When expressed in cultured mammalian cells, full-length mVps24p is cytosolic. However, when cells expressing the full-length mVps24p are co-transfected with a mutated form of mVps4p (which is defective in ATP hydrolysis), or when a N-terminal construct of mVps24p is expressed, the class-E cellular phenotype with swollen vacuoles is induced and mVps24p is membrane-associated. Furthermore, the accumulation of the N-terminal mVps24p construct on the swollen endosomal membranes is abrogated when phosphatidylinositol 3,5-bisphosphate synthesis is blocked with wortmannin. These data provide the first direct link between phosphatidylinositol 3,5-bisphosphate and the protein machinery involved in the production of the class-E cellular phenotype. We hypothesize that accumulation of Vps24 on membranes occurs when membrane association (dependent on interaction of phosphatidylinositol 3,5-bisphosphate with the N-terminal domain of the protein) is uncoupled from membrane disassociation (driven by Vps4p).

Received for publication, June 27, 2003

^* This work was supported by the Medical Research Council UK and Diabetes UK (to G. D. H.) and the Wellcome Trust (Program Grant 060554 to B. V. L. P.). 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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AY150169.

To whom correspondence should be addressed. Tel.: 44-1-225-384278; Fax: 44-1-225-386779; E-mail: bssprw{at}bath.ac.uk.

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

This article has been cited by other articles:

				J. van der Zee, H. Urwin, S. Engelborghs, M. Bruyland, R. Vandenberghe, B. Dermaut, T. De Pooter, K. Peeters, P. Santens, P. P. De Deyn, et al. CHMP2B C-truncating mutations in frontotemporal lobar degeneration are associated with an aberrant endosomal phenotype in vitro Hum. Mol. Genet., January 15, 2008; 17(2): 313 - 322. [Abstract] [Full Text] [PDF]

				J. A. Efe, R. J. Botelho, and S. D. Emr Atg18 Regulates Organelle Morphology and Fab1 Kinase Activity Independent of Its Membrane Recruitment by Phosphatidylinositol 3,5-Bisphosphate Mol. Biol. Cell, November 1, 2007; 18(11): 4232 - 4244. [Abstract] [Full Text] [PDF]

				P. E. Row, H. Liu, S. Hayes, R. Welchman, P. Charalabous, K. Hofmann, M. J. Clague, C. M. Sanderson, and S. Urbe The MIT Domain of UBPY Constitutes a CHMP Binding and Endosomal Localization Signal Required for Efficient Epidermal Growth Factor Receptor Degradation J. Biol. Chem., October 19, 2007; 282(42): 30929 - 30937. [Abstract] [Full Text] [PDF]

				Y. M. Ma, E. Boucrot, J. Villen, E. B. Affar, S. P. Gygi, H. G. Gottlinger, and T. Kirchhausen Targeting of AMSH to Endosomes Is Required for Epidermal Growth Factor Receptor Degradation J. Biol. Chem., March 30, 2007; 282(13): 9805 - 9812. [Abstract] [Full Text] [PDF]

				A. Zamborlini, Y. Usami, S. R. Radoshitzky, E. Popova, G. Palu, and H. Gottlinger Release of autoinhibition converts ESCRT-III components into potent inhibitors of HIV-1 budding PNAS, December 12, 2006; 103(50): 19140 - 19145. [Abstract] [Full Text] [PDF]

				A. C. Rutherford, C. Traer, T. Wassmer, K. Pattni, M. V. Bujny, J. G. Carlton, H. Stenmark, and P. J. Cullen The mammalian phosphatidylinositol 3-phosphate 5-kinase (PIKfyve) regulates endosome-to-TGN retrograde transport J. Cell Sci., October 1, 2006; 119(19): 3944 - 3957. [Abstract] [Full Text] [PDF]

				W. S. A. Maaty, A. C. Ortmann, M. Dlakic, K. Schulstad, J. K. Hilmer, L. Liepold, B. Weidenheft, R. Khayat, T. Douglas, M. J. Young, et al. Characterization of the Archaeal Thermophile Sulfolobus Turreted Icosahedral Virus Validates an Evolutionary Link among Double-Stranded DNA Viruses from All Domains of Life. J. Virol., August 1, 2006; 80(15): 7625 - 7635. [Abstract] [Full Text] [PDF]

				O. C. Ikonomov, D. Sbrissa, and A. Shisheva Localized PtdIns 3,5-P2 synthesis to regulate early endosome dynamics and fusion Am J Physiol Cell Physiol, August 1, 2006; 291(2): C393 - C404. [Abstract] [Full Text] [PDF]

				A.-S. Nicot, H. Fares, B. Payrastre, A. D. Chisholm, M. Labouesse, and J. Laporte The Phosphoinositide Kinase PIKfyve/Fab1p Regulates Terminal Lysosome Maturation in Caenorhabditis elegans Mol. Biol. Cell, July 1, 2006; 17(7): 3062 - 3074. [Abstract] [Full Text] [PDF]

				E. E. Johnson, J. H. Overmeyer, W. T. Gunning, and W. A. Maltese Gene silencing reveals a specific function of hVps34 phosphatidylinositol 3-kinase in late versus early endosomes J. Cell Sci., April 1, 2006; 119(7): 1219 - 1232. [Abstract] [Full Text] [PDF]

				K. Bowers, S. C. Piper, M. A. Edeling, S. R. Gray, D. J. Owen, P. J. Lehner, and J. P. Luzio Degradation of Endocytosed Epidermal Growth Factor and Virally Ubiquitinated Major Histocompatibility Complex Class I Is Independent of Mammalian ESCRTII J. Biol. Chem., February 24, 2006; 281(8): 5094 - 5105. [Abstract] [Full Text] [PDF]

				K. Lindmo and H. Stenmark Regulation of membrane traffic by phosphoinositide 3-kinases J. Cell Sci., February 15, 2006; 119(4): 605 - 614. [Abstract] [Full Text] [PDF]

				P. Berger, I. Berger, C. Schaffitzel, K. Tersar, B. Volkmer, and U. Suter Multi-level regulation of myotubularin-related protein-2 phosphatase activity by myotubularin-related protein-13/set-binding factor-2 Hum. Mol. Genet., February 15, 2006; 15(4): 569 - 579. [Abstract] [Full Text] [PDF]

				C. M. Sanderson, J. W. Connell, T. L. Edwards, N. A. Bright, S. Duley, A. Thompson, J. P. Luzio, and E. Reid Spastin and atlastin, two proteins mutated in autosomal-dominant hereditary spastic paraplegia, are binding partners Hum. Mol. Genet., January 15, 2006; 15(2): 307 - 318. [Abstract] [Full Text] [PDF]

				Y. Ling, A. J. Smith, and G. T. Morgan A sequence motif conserved in diverse nuclear proteins identifies a protein interaction domain utilised for nuclear targeting by human TFIIS. Nucleic Acids Res., January 1, 2006; 34(8): 2219 - 2229. [Abstract] [Full Text] [PDF]

				Y. Lin, L. A. Kimpler, T. V. Naismith, J. M. Lauer, and P. I. Hanson Interaction of the Mammalian Endosomal Sorting Complex Required for Transport (ESCRT) III Protein hSnf7-1 with Itself, Membranes, and the AAA+ ATPase SKD1 J. Biol. Chem., April 1, 2005; 280(13): 12799 - 12809. [Abstract] [Full Text] [PDF]

				D. Sbrissa and A. Shisheva Acquisition of Unprecedented Phosphatidylinositol 3,5-Bisphosphate Rise in Hyperosmotically Stressed 3T3-L1 Adipocytes, Mediated by ArPIKfyve-PIKfyve Pathway J. Biol. Chem., March 4, 2005; 280(9): 7883 - 7889. [Abstract] [Full Text] [PDF]

				D. C. Berwick, G. C. Dell, G. I. Welsh, K. J. Heesom, I. Hers, L. M. Fletcher, F. T. Cooke, and J. M. Tavare Protein kinase B phosphorylation of PIKfyve regulates the trafficking of GLUT4 vesicles J. Cell Sci., December 1, 2004; 117(25): 5985 - 5993. [Abstract] [Full Text] [PDF]

				D. J. Katzmann, S. Sarkar, T. Chu, A. Audhya, and S. D. Emr Multivesicular Body Sorting: Ubiquitin Ligase Rsp5 Is Required for the Modification and Sorting of Carboxypeptidase S Mol. Biol. Cell, February 1, 2004; 15(2): 468 - 480. [Abstract] [Full Text] [PDF]