Arrestin 3 mediates endocytosis of CCR7 following ligation of CCL19 but not CCL21.
Internalization of ligand bound G protein-coupled receptors, an important cellular function that mediates receptor desensitization, takes place via distinct pathways, which are often unique for each receptor. The C-C chemokine receptor (CCR7) G protein-coupled receptor is expressed on naive T cells, dendritic cells, and NK cells and has two endogenous ligands, CCL19 and CCL21. Following binding of CCL21, 21 +/- 4% of CCR7 is internalized in the HuT 78 human T cell lymphoma line, while 76 +/- 8% of CCR7 is internalized upon binding to CCL19. To determine whether arrestins mediated differential internalization of CCR7/CCL19 vs CCR7/CCL21, we used small interfering RNA (siRNA) to knock down expression of arrestin 2 or arrestin 3 in HuT 78 cells. Independent of arrestin 2 or arrestin 3 expression, CCR7/CCL21 internalized. In contrast, following depletion of arrestin 3, CCR7/CCL19 failed to internalize. To examine the consequence of complete loss of both arrestin 2 and arrestin 3 on CCL19/CCR7 internalization, we examined CCR7 internalization in arrestin 2(-/-)/arrestin 3(-/-) murine embryonic fibroblasts. Only reconstitution with arrestin 3-GFP but not arrestin 2-GFP rescued internalization of CCR7/CCL19. Loss of arrestin 2 or arrestin 3 blocked migration to CCL19 but had no effect on migration to CCL21. Using immunofluorescence microscopy, we found that arrestins do not cluster at the membrane with CCR7 following ligand binding but cap with CCR7 during receptor internalization. These are the first studies that define a role for arrestin 3 in the internalization of a chemokine receptor following binding of one but not both endogenous ligands.
Byers
Melissa A
MA
Department of Microbiology, Molecular Genetics and Immunology, The University of Kansas Medical Center, Kansas City, KS 66160, USA.
Calloway
Psachal A
PA
Shannon
Laurie
L
Cunningham
Heather D
HD
Smith
Sarah
S
Li
Fang
F
Fassold
Brian C
BC
Vines
Charlotte M
CM
D000818:Animals; D019390:Arrestins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D054415:Chemokine CCL19 / Q000378:metabolism*; D054421:Chemokine CCL21 / Q000378:metabolism; D004705:Endocytosis / Q000276:immunology*; D006801:Humans; D019169:Jurkat Cells; D008024:Ligands; D051379:Mice; D011485:Protein Binding / Q000276:immunology; D034741:RNA, Small Interfering / Q000502:physiology; D011940:Receptor Aggregation / Q000276:immunology; D054400:Receptors, CCR7 / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D000071557:beta-Arrestins
D019390:Arrestins; C515460:CCL19 protein, human; C515478:CCL21 protein, human; C515370:CCR7 protein, human; D054415:Chemokine CCL19; D054421:Chemokine CCL21; D008024:Ligands; D034741:RNA, Small Interfering; D054400:Receptors, CCR7; C082333:arrestin3; D000071557:beta-Arrestins
Mol Biol Med. 1987 Dec;4(6):385-96
3501821
J Biol Chem. 1997 Jun 6;272(23):15011-6
9169476
Int Immunol. 1998 Jul;10(7):901-10
9701028
Cell Immunol. 1999 May 1;193(2):226-35
10222066
J Biol Chem. 2002 Dec 20;277(51):49212-9
12370187
J Cell Biol. 1994 May;125(3):639-49
7513706
Mol Pharmacol. 2003 Jan;63(1):9-18
12488531
FEMS Microbiol Lett. 2004 Jul 1;236(1):1-6
15212783
Can J Physiol Pharmacol. 1996 Oct;74(10):1095-110
9022829
Science. 1999 Jan 29;283(5402):655-61
9924018
J Biol Chem. 2004 May 28;279(22):23214-22
15054093
Science. 1996 Jan 19;271(5247):363-6
8553074
J Biol Chem. 2004 Sep 24;279(39):40276-82
15231820
J Exp Med. 1973 Feb 1;137(2):387-410
4568301
Proc Natl Acad Sci U S A. 2004 Feb 10;101(6):1548-53
14757828
Biochem Soc Trans. 1996 Nov;24(4):953-9
8968491
J Biol Chem. 2006 Jan 13;281(2):1261-73
16280323
J Clin Pathol. 2005 Oct;58(10):1057-63
16189151
Nat Immunol. 2005 Sep;6(9):889-94
16116468
Biochemistry. 2001 Mar 27;40(12):3467-75
11297412
J Immunol. 2006 Aug 15;177(4):2314-23
16887992
J Biol Chem. 2000 Apr 14;275(15):11312-9
10753943
Pharmacol Rev. 2001 Mar;53(1):1-24
11171937
Annu Rev Pharmacol Toxicol. 2008;48:601-29
17995450
Cell Mol Life Sci. 2002 Dec;59(12):2172-83
12568343
J Biol Chem. 1997 Nov 21;272(47):29426-9
9367998
J Immunol. 1997 Dec 1;159(11):5671-9
9548511
Eur J Immunol. 1998 Jun;28(6):2025-34
9645384
Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3712-7
10097102
J Biol Chem. 2005 Apr 22;280(16):16076-87
15708858
Nature. 1996 Oct 3;383(6599):447-50
8837779
Blood. 1999 May 15;93(10):3225-32
10233873
Arthritis Rheum. 2003 Nov;48(11):3187-201
14613282
J Immunol Methods. 1980;33(3):239-47
6989919
J Immunol. 1989 Jul 1;143(1):229-38
2732469
Proc Natl Acad Sci U S A. 2003 Sep 16;100(19):10782-7
12949261
J Cell Biol. 1998 May 18;141(4):1053-9
9585422
J Immunol. 2007 Nov 15;179(10):6485-93
17982037
Immunity. 2001 Oct;15(4):507-19
11672534
J Leukoc Biol. 1997 Nov;62(5):634-44
9365118
J Biol Chem. 1999 Apr 16;274(16):10677-80
10196135
Arthritis Rheum. 2005 Jun;52(6):1773-84
15934082
J Biol Chem. 2003 Nov 14;278(46):45954-9
12960162
J Biol Chem. 2002 Mar 15;277(11):8970-8
11777932
Eur J Immunol. 2001 Nov;31(11):3291-7
11745346
Neuroreport. 2006 Feb 6;17(2):173-7
16407766
Nat Immunol. 2005 Sep;6(9):895-901
16116469
J Immunol. 2002 Jul 1;169(1):424-33
12077273
J Biol Chem. 1993 Oct 25;268(30):22273-6
8226735
Mol Endocrinol. 2000 Dec;14(12):2040-53
11117533
J Immunol. 1998 Sep 15;161(6):3096-102
9743376
Biochemistry. 1999 Feb 23;38(8):2240-7
10029516
J Clin Invest. 1981 Feb;67(2):523-30
6970206
Semin Immunol. 2003 Oct;15(5):271-6
15001176
Immunity. 2002 Feb;16(2):205-18
11869682
J Biol Chem. 2001 Nov 23;276(47):43663-7
11546805
Pharmacol Ther. 1997 Aug;75(2):135-51
9428002
FEBS Lett. 1992 Apr 27;301(3):310-4
1577171
J Immunol. 1999 May 15;162(10):5981-5
10229836
Cell Immunol. 2000 Jan 10;199(1):8-14
10675270
J Mol Neurosci. 2005;26(2-3):253-64
16012199
Biochem Biophys Res Commun. 1999 Oct 5;263(3):685-90
10512740
J Biol Chem. 2004 Jun 4;279(23):24578-84
15051714
Microcirculation. 2004 Sep;11(6):493-502
15371130
Immunol Rev. 2002 Aug;186:37-46
12234360
J Biol Chem. 2003 Feb 7;278(6):4041-7
12424254
J Biol Chem. 2003 Oct 24;278(43):41581-4
12947104
J Biol Chem. 1996 Aug 2;271(31):18302-5
8702465
Eur J Immunol. 2007 Feb;37(2):351-7
17236235
Curr Drug Targets. 2007 Jan;8(1):137-46
17266537
Curr Opin Cell Biol. 1999 Apr;11(2):177-83
10209148
J Biol Chem. 2004 Oct 22;279(43):45175-84
15302864
Nat Methods. 2006 Sep;3(9):670-6
16929310
Proc Natl Acad Sci U S A. 1998 Jan 6;95(1):258-63
9419363
Mol Cell Biol. 2004 Oct;24(20):8929-37
15456867
Blood. 2000 Jan 1;95(1):30-8
10607681
Neuroreport. 2001 Oct 8;12(14):3159-63
11568656
Gut. 2006 Feb;55(2):220-7
16118351
J Exp Med. 2001 Jan 15;193(2):207-18
11148224
Science. 2001 Nov 9;294(5545):1307-13
11588219
Science. 1998 Jan 16;279(5349):381-4
9430588
Circ Res. 2006 Sep 15;99(6):570-82
16973913