Filamin a binds to CCR2B and regulates its internalization.
The chemokine (C-C motif) receptor 2B (CCR2B) is one of the two isoforms of the receptor for monocyte chemoattractant protein-1 (CCL2), the major chemoattractant for monocytes, involved in an array of chronic inflammatory diseases. Employing the yeast two-hybrid system, we identified the actin-binding protein filamin A (FLNa) as a protein that associates with the carboxyl-terminal tail of CCR2B. Co-immunoprecipitation experiments and in vitro pull down assays demonstrated that FLNa binds constitutively to CCR2B. The colocalization of endogenous CCR2B and filamin A was detected at the surface and in internalized vesicles of THP-1 cells. In addition, CCR2B and FLNa were colocalized in lamellipodia structures of CCR2B-expressing A7 cells. Expression of the receptor in filamin-deficient M2 cells together with siRNA experiments knocking down FLNa in HEK293 cells, demonstrated that lack of FLNa delays the internalization of the receptor. Furthermore, depletion of FLNa in THP-1 monocytes by RNA interference reduced the migration of cells in response to MCP-1. Therefore, FLNa emerges as an important protein for controlling the internalization and spatial localization of the CCR2B receptor in different dynamic membrane structures.
Minsaas
Laura
L
Department of Biomedicine, University of Bergen, Bergen, Norway.
Planagumà
Jesús
J
Madziva
Michael
M
Krakstad
Beate F
BF
Masià-Balagué
Míriam
M
Katz
Arieh A
AA
Aragay
Anna M
AM
D000199:Actins / Q000378:metabolism; D000818:Animals; D019390:Arrestins / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D018932:Chemokine CCL2 / Q000494:pharmacology; D003285:Contractile Proteins / Q000378:metabolism*; D064448:Filamins; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism*; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D054390:Receptors, CCR2 / Q000737:chemistry / Q000378:metabolism*; D000071557:beta-Arrestins
D000199:Actins; D019390:Arrestins; D018932:Chemokine CCL2; D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins; D054390:Receptors, CCR2; D000071557:beta-Arrestins
Proc Natl Acad Sci U S A. 2008 Jul 22;105(29):10101-6
18632580
Biochem Soc Trans. 2009 Oct;37(Pt 5):1072-6
19754454
Nat Immunol. 2008 Sep;9(9):953-9
18711432
Nat Immunol. 2008 Sep;9(9):949-52
18711431
J Biol Chem. 2001 Sep 14;276(37):34871-9
11390379
J Leukoc Biol. 2006 Dec;80(6):1214-21
16943386
AIDS Res Hum Retroviruses. 2008 Dec;24(12):1527-36
19018669
Nat Cell Biol. 2007 Jul;9(7):838-46
17572668
J Biol Chem. 2003 Mar 21;278(12):10408-16
12531889
J Biol Chem. 2001 Sep 14;276(37):34880-7
11390380
Biochem J. 2008 Feb 1;409(3):635-49
18177271
J Biol Chem. 2005 Mar 25;280(12):11140-6
15657061
Trends Biochem Sci. 2006 Jul;31(7):411-9
16781869
Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5258-63
11320256
J Biol Chem. 2005 Apr 1;280(13):12774-80
15687500
Nat Cell Biol. 2002 Sep;4(9):691-8
12198492
Nat Rev Mol Cell Biol. 2001 Feb;2(2):138-45
11252955
Annu Rev Pharmacol Toxicol. 1998;38:289-319
9597157
J Cell Biol. 1997 Dec 29;139(7):1719-33
9412467
FEBS Lett. 2002 Mar 13;514(2-3):184-8
11943148
Mol Endocrinol. 2007 Sep;21(9):2242-54
17536008
Mol Cell Biol. 2006 May;26(9):3432-45
16611986
FEBS Lett. 2006 Mar 20;580(7):1795-800
16513120
Mol Pharmacol. 2003 Sep;64(3):773-82
12920215
Pharmacology. 2002 Dec;66(4):173-81
12393939
Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2122-8
10051605
J Cell Biol. 1998 Sep 7;142(5):1245-56
9732285
Mol Pharmacol. 2003 Nov;64(5):1092-100
14573758
J Immunol. 1996 Dec 15;157(12):5606-12
8955213
Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2985-90
9501202
J Biol Chem. 2009 Dec 11;284(50):35113-21
19828450
Cell. 2005 Oct 21;123(2):305-20
16239147
J Leukoc Biol. 2004 May;75(5):901-9
14761932
Nat Immunol. 2005 Aug;6(8):827-35
15995708
J Neurosci. 2000 Dec 1;20(23):8736-44
11102480
J Cell Biol. 2001 Nov 12;155(4):511-7
11706047
Genome Biol. 2006;7(10):R100
17076895
Biochem Pharmacol. 2002 Mar 1;63(5):859-63
11911837