Endosomal-sorting Complexes Required for Transport (ESCRT) Pathway-dependent Endosomal Traffic Regulates the Localization of Active Src at Focal Adhesions

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2010 Sep 1

PMID 20805499

Citations 39

Authors

Chun Tu

Cesar F Ortega-Cava

Paul Winograd

Marissa Jo Stanton

Alagarsamy Lakku Reddi

Ingrid Dodge

Ranjana Arya

Manjari Dimri

Robert J Clubb

Mayumi Naramura

Kay-Uwe Wagner

Vimla Band

Hamid Band

Affiliations

Soon will be listed here.

Abstract

Active Src localization at focal adhesions (FAs) is essential for cell migration. How this pool is linked mechanistically to the large pool of Src at late endosomes (LEs)/lysosomes (LY) is not well understood. Here, we used inducible Tsg101 gene deletion, TSG101 knockdown, and dominant-negative VPS4 expression to demonstrate that the localization of activated cellular Src and viral Src at FAs requires the endosomal-sorting complexes required for transport (ESCRT) pathway. Tsg101 deletion also led to impaired Src-dependent activation of STAT3 and focal adhesion kinase and reduced cell migration. Impairment of the ESCRT pathway or Rab7 function led to the accumulation of active Src at aberrant LE/LY compartments followed by its loss. Analyses using fluorescence recovery after photo-bleaching show that dynamic mobility of Src in endosomes is ESCRT pathway-dependent. These results reveal a critical role for an ESCRT pathway-dependent LE/LY trafficking step in Src function by promoting localization of active Src to FAs.

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References

Ichioka F, Takaya E, Suzuki H, Kajigaya S, Buchman V, Shibata H . HD-PTP and Alix share some membrane-traffic related proteins that interact with their Bro1 domains or proline-rich regions. Arch Biochem Biophys. 2006; 457(2):142-9. DOI: 10.1016/j.abb.2006.11.008. View

Garrus J, von Schwedler U, Pornillos O, Morham S, Zavitz K, Wang H . Tsg101 and the vacuolar protein sorting pathway are essential for HIV-1 budding. Cell. 2001; 107(1):55-65. DOI: 10.1016/s0092-8674(01)00506-2. View

Oneyama C, Iino T, Saito K, Suzuki K, Ogawa A, Okada M . Transforming potential of Src family kinases is limited by the cholesterol-enriched membrane microdomain. Mol Cell Biol. 2009; 29(24):6462-72. PMC: 2786866. DOI: 10.1128/MCB.00941-09. View

Morita E, Sandrin V, Chung H, Morham S, Gygi S, Rodesch C . Human ESCRT and ALIX proteins interact with proteins of the midbody and function in cytokinesis. EMBO J. 2007; 26(19):4215-27. PMC: 2230844. DOI: 10.1038/sj.emboj.7601850. View

Klinghoffer R, Sachsenmaier C, Cooper J, Soriano P . Src family kinases are required for integrin but not PDGFR signal transduction. EMBO J. 1999; 18(9):2459-71. PMC: 1171328. DOI: 10.1093/emboj/18.9.2459. View

Schmidt M, Dikic I, Bogler O . Src phosphorylation of Alix/AIP1 modulates its interaction with binding partners and antagonizes its activities. J Biol Chem. 2004; 280(5):3414-25. PMC: 1180480. DOI: 10.1074/jbc.M409839200. View

Goodwin J, Kenworthy A . Photobleaching approaches to investigate diffusional mobility and trafficking of Ras in living cells. Methods. 2005; 37(2):154-64. DOI: 10.1016/j.ymeth.2005.05.013. View

Babst M, Odorizzi G, Estepa E, Emr S . Mammalian tumor susceptibility gene 101 (TSG101) and the yeast homologue, Vps23p, both function in late endosomal trafficking. Traffic. 2001; 1(3):248-58. DOI: 10.1034/j.1600-0854.2000.010307.x. View

Thien C, Langdon W . Negative regulation of PTK signalling by Cbl proteins. Growth Factors. 2005; 23(2):161-7. DOI: 10.1080/08977190500153763. View

10.

Sato I, Obata Y, Kasahara K, Nakayama Y, Fukumoto Y, Yamasaki T . Differential trafficking of Src, Lyn, Yes and Fyn is specified by the state of palmitoylation in the SH4 domain. J Cell Sci. 2009; 122(Pt 7):965-75. DOI: 10.1242/jcs.034843. View

11.

Turkson J, Bowman T, Garcia R, Caldenhoven E, de Groot R, Jove R . Stat3 activation by Src induces specific gene regulation and is required for cell transformation. Mol Cell Biol. 1998; 18(5):2545-52. PMC: 110634. DOI: 10.1128/MCB.18.5.2545. View

12.

Babst M, Wendland B, Estepa E, Emr S . The Vps4p AAA ATPase regulates membrane association of a Vps protein complex required for normal endosome function. EMBO J. 1998; 17(11):2982-93. PMC: 1170638. DOI: 10.1093/emboj/17.11.2982. View

13.

Fujita H, Yamanaka M, Imamura K, Tanaka Y, Nara A, Yoshimori T . A dominant negative form of the AAA ATPase SKD1/VPS4 impairs membrane trafficking out of endosomal/lysosomal compartments: class E vps phenotype in mammalian cells. J Cell Sci. 2002; 116(Pt 2):401-14. DOI: 10.1242/jcs.00213. View

14.

Donepudi M, Resh M . c-Src trafficking and co-localization with the EGF receptor promotes EGF ligand-independent EGF receptor activation and signaling. Cell Signal. 2008; 20(7):1359-67. PMC: 2459337. DOI: 10.1016/j.cellsig.2008.03.007. View

15.

Yorikawa C, Shibata H, Waguri S, Hatta K, Horii M, Katoh K . Human CHMP6, a myristoylated ESCRT-III protein, interacts directly with an ESCRT-II component EAP20 and regulates endosomal cargo sorting. Biochem J. 2004; 387(Pt 1):17-26. PMC: 1134928. DOI: 10.1042/BJ20041227. View

16.

Scott M, Zappacosta F, Kim E, Annan R, Miller W . Identification of novel SH3 domain ligands for the Src family kinase Hck. Wiskott-Aldrich syndrome protein (WASP), WASP-interacting protein (WIP), and ELMO1. J Biol Chem. 2002; 277(31):28238-46. DOI: 10.1074/jbc.M202783200. View

17.

Yoshimori T, YAMAGATA F, Yamamoto A, Mizushima N, Kabeya Y, Nara A . The mouse SKD1, a homologue of yeast Vps4p, is required for normal endosomal trafficking and morphology in mammalian cells. Mol Biol Cell. 2000; 11(2):747-63. PMC: 14807. DOI: 10.1091/mbc.11.2.747. View

18.

Kasahara K, Nakayama Y, Kihara A, Matsuda D, Ikeda K, Kuga T . Rapid trafficking of c-Src, a non-palmitoylated Src-family kinase, between the plasma membrane and late endosomes/lysosomes. Exp Cell Res. 2007; 313(12):2651-66. DOI: 10.1016/j.yexcr.2007.05.001. View

19.

Ruland J, Sirard C, Elia A, Macpherson D, Wakeham A, Li L . p53 accumulation, defective cell proliferation, and early embryonic lethality in mice lacking tsg101. Proc Natl Acad Sci U S A. 2001; 98(4):1859-64. PMC: 29347. DOI: 10.1073/pnas.98.4.1859. View

20.

Mariotti M, Castiglioni S, Garcia-Manteiga J, Beguinot L, Maier J . HD-PTP inhibits endothelial migration through its interaction with Src. Int J Biochem Cell Biol. 2008; 41(3):687-93. DOI: 10.1016/j.biocel.2008.08.005. View