Endosomal-sorting complexes required for transport (ESCRT) pathway-dependent endosomal traffic regulates the localization of active Src at focal adhesions.
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.
Tu
Chun
C
Eppley Institute for Research in Cancer and Allied Diseases and University of Nebraska Medical Center-Eppley Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Ortega-Cava
Cesar F
CF
Winograd
Paul
P
Stanton
Marissa Jo
MJ
Reddi
Alagarsamy Lakku
AL
Dodge
Ingrid
I
Arya
Ranjana
R
Dimri
Manjari
M
Clubb
Robert J
RJ
Naramura
Mayumi
M
Wagner
Kay-Uwe
KU
Band
Vimla
V
Band
Hamid
H
D000818:Animals; D002448:Cell Adhesion; D002465:Cell Movement; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism*; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D051379:Mice; D021381:Protein Transport; D014157:Transcription Factors / Q000378:metabolism; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism*; D000099177:Tsg101 Protein
D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D014157:Transcription Factors; D019061:src-Family Kinases; D000099177:Tsg101 Protein
Arch Biochem Biophys. 2007 Jan 15;457(2):142-9
17174262
Cell. 2001 Oct 5;107(1):55-65
11595185
Mol Cell Biol. 2009 Dec;29(24):6462-72
19822664
EMBO J. 2007 Oct 3;26(19):4215-27
17853893
EMBO J. 1999 May 4;18(9):2459-71
10228160
J Biol Chem. 2005 Feb 4;280(5):3414-25
15557335
Methods. 2005 Oct;37(2):154-64
16288889
Traffic. 2000 Mar;1(3):248-58
11208108
Growth Factors. 2005 Jun;23(2):161-7
16019438
J Cell Sci. 2009 Apr 1;122(Pt 7):965-75
19258394
Mol Cell Biol. 1998 May;18(5):2545-52
9566874
EMBO J. 1998 Jun 1;17(11):2982-93
9606181
J Cell Sci. 2003 Jan 15;116(Pt 2):401-14
12482925
Cell Signal. 2008 Jul;20(7):1359-67
18448311
Biochem J. 2005 Apr 1;387(Pt 1):17-26
15511219
J Biol Chem. 2002 Aug 2;277(31):28238-46
12029088
Mol Biol Cell. 2000 Feb;11(2):747-63
10679028
Exp Cell Res. 2007 Jul 15;313(12):2651-66
17537435
Proc Natl Acad Sci U S A. 2001 Feb 13;98(4):1859-64
11172041
Int J Biochem Cell Biol. 2009 Mar;41(3):687-93
18762272
J Biol Chem. 2005 Feb 18;280(7):6036-46
15579470
J Cell Sci. 2005 Jul 15;118(Pt 14):3003-17
16014378
Biochemistry. 2009 Oct 13;48(40):9321-3
19673488
Immunity. 2010 Apr 23;32(4):531-40
20399684
Biochem J. 2005 Nov 1;391(Pt 3):677-85
16004603
J Biol Chem. 2005 Mar 11;280(10):9258-64
15611048
Mol Cell. 1999 May;3(5):629-38
10360179
J Cell Sci. 2007 Aug 1;120(Pt 15):2555-64
17623777
Trends Cell Biol. 2008 Jul;18(7):322-9
18515107
Mol Cell Biol. 1996 Oct;16(10):5623-33
8816475
Biochem J. 2002 Jul 15;365(Pt 2):349-53
11931639
J Biol Chem. 2001 Apr 13;276(15):11735-42
11134028
Trends Cell Biol. 2006 Jun;16(6):317-26
16716591
Cancer Cell. 2004 Sep;6(3):209-14
15380511
Mol Biol Cell. 2000 Jan;11(1):227-39
10637304
Mol Cell Biol. 2003 Jan;23(1):150-62
12482969
J Cell Biol. 1996 Dec;135(6 Pt 1):1551-64
8978822
Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7626-31
12802020
Oncogene. 2004 Oct 18;23(48):7906-9
15489908
Mol Cell. 2008 May 23;30(4):426-36
18498747
Nature. 2007 Oct 11;449(7163):740-4
17928862
J Cell Biol. 1998 Mar 9;140(5):1075-89
9490721
Science. 2007 Jun 29;316(5833):1908-12
17556548
Mol Cell Biol. 2006 Mar;26(5):1932-47
16479011
Proc Natl Acad Sci U S A. 2008 Apr 29;105(17):6308-13
18434552
Dev Cell. 2004 Dec;7(6):855-69
15572128
J Cell Biol. 2009 Apr 20;185(2):357-70
19364917
Cell. 1987 Apr 10;49(1):65-73
3103925
EMBO J. 1998 Jan 2;17(1):81-92
9427743
J Biol Chem. 2002 Nov 8;277(45):43216-23
12205095
J Virol. 1986 Dec;60(3):840-8
3023673