Sorting motifs of the endosomal/lysosomal CLC chloride transporters.
The CLC protein family contains plasma membrane chloride channels and the intracellular chloride-proton exchangers ClC-3-7. The latter proteins mainly reside on the various compartments of the endosomal-lysosomal system where they are involved in the luminal acidification or chloride accumulation. Although their partially overlapping subcellular distribution has been studied extensively, little is known about their targeting mechanism. In a comprehensive study we now performed pulldown experiments to systematically map the differential binding of adaptor proteins of the endosomal sorting machinery (adaptor proteins and GGAs (Golgi-localized, γ-ear containing, Arf binding)) as well as clathrin to the cytosolic regions of the intracellular CLCs. The resulting interaction pattern fitted well to the known subcellular localizations of the CLCs. By mutating potential sorting motifs, we could locate almost all binding sites, including one already known for ClC-3 and several new motifs for ClC-5, -6, and -7. The impact of the identified binding sites on the subcellular localization of CLC transporters was determined by heterologous expression of mutants. Surprisingly, some vesicular CLCs retained their localization after disruption of interaction sites. However, ClC-7 could be partially shifted from lysosomes to the plasma membrane by combined mutation of N-terminal sorting motifs. The localization of its β-subunit, Ostm1, was determined by that of ClC-7. Ostm1 was not capable of redirecting ClC-7 to lysosomes.
Stauber
Tobias
T
Leibniz-Institut für Molekulare Pharmakologie and Max-Delbrück-Centrum für Molekulare Medizin, Berlin D-13125, Germany.
Jentsch
Thomas J
TJ
D020816:Amino Acid Motifs; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D018118:Chloride Channels / Q000235:genetics / Q000378:metabolism*; D002966:Clathrin / Q000235:genetics / Q000378:metabolism; D011992:Endosomes / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D008247:Lysosomes / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D010449:Peptide Mapping; D021382:Protein Sorting Signals / Q000502:physiology*; D021381:Protein Transport / Q000502:physiology; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism; D014981:Xenopus
D018118:Chloride Channels; D002966:Clathrin; D008565:Membrane Proteins; C489946:OSTM1 protein, human; D021382:Protein Sorting Signals; D044767:Ubiquitin-Protein Ligases
FASEB J. 2002 Jun;16(8):863-5
11967229
J Biol Chem. 2005 Jan 14;280(2):1241-7
15504734
Circ Res. 2007 Sep 28;101(7):663-71
17673675
Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):13362-6
8917596
Crit Rev Biochem Mol Biol. 2008 Jan-Feb;43(1):3-36
18307107
Annu Rev Biochem. 2003;72:395-447
12651740
J Biol Chem. 1995 Dec 29;270(52):31172-7
8537381
J Biol Chem. 2002 Jan 4;277(1):566-74
11675385
J Mol Med (Berl). 1997 Mar;75(3):160-3
9106071
Biochim Biophys Acta. 2009 Apr;1793(4):605-14
19046998
Proc Natl Acad Sci U S A. 2006 Sep 12;103(37):13854-9
16950870
J Cell Physiol. 2006 Mar;206(3):792-8
16222710
Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):8075-80
9653142
J Biol Chem. 2004 Dec 31;279(53):54996-5007
15489223
Nat Rev Mol Cell Biol. 2004 Jan;5(1):23-32
14708007
J Biol Chem. 2001 Jan 26;276(4):2347-53
11035003
Nature. 2006 Mar 9;440(7081):220-3
16525474
Trends Cell Biol. 2004 Apr;14(4):167-74
15066634
J Neurosci. 2008 Oct 15;28(42):10587-98
18923035
J Biol Chem. 2010 Jun 4;285(23):17595-603
20351103
J Biol Chem. 2003 Aug 1;278(31):29267-77
12746443
J Cell Sci. 2006 May 15;119(Pt 10):2003-14
16621890
J Biol Chem. 2010 Jul 9;285(28):21689-97
20466723
Neuron. 1999 Mar;22(3):537-48
10197533
Nature. 2001 Nov 29;414(6863):558-61
11734858
Am J Physiol Renal Physiol. 2005 Oct;289(4):F850-62
15942052
J Biol Chem. 2007 Sep 28;282(39):29022-29031
17652080
Am J Physiol Cell Physiol. 2002 Jun;282(6):C1483-91
11997263
Nature. 2002 Jan 17;415(6869):287-94
11796999
Science. 2010 Jun 11;328(5984):1398-401
20430975
Neuron. 2001 Jan;29(1):185-96
11182090
Am J Physiol Cell Physiol. 2001 Feb;280(2):C373-81
11208533
J Physiol. 2007 Feb 1;578(Pt 3):633-40
17110406
Pflugers Arch. 2003 Jan;445(4):456-62
12548389
Trends Biochem Sci. 1997 Jan;22(1):12-3
9020585
Am J Physiol. 1999 Dec;277(6):F957-65
10600943
Cell. 2001 Jan 26;104(2):205-15
11207362
EMBO J. 2008 Nov 5;27(21):2907-17
18833191
Science. 2010 Jun 11;328(5984):1401-3
20430974
J Cell Physiol. 2009 Dec;221(3):650-7
19711355
EMBO J. 2005 Mar 9;24(5):1079-91
15706348
FASEB J. 2009 Dec;23(12):4056-68
19661288
J Cell Sci. 2005 Sep 15;118(Pt 18):4243-52
16155254
Mol Cancer Ther. 2007 Mar;6(3):979-86
17363491
FEBS Lett. 1995 Dec 11;377(1):15-20
8543009
EMBO J. 1997 Nov 3;16(21):6325-36
9351815
FASEB J. 2006 Nov;20(13):2390-2
17023393
Traffic. 2009 Nov;10(11):1696-710
19847956
J Biol Chem. 2003 Feb 21;278(8):6440-9
12471024
PLoS One. 2007 May 30;2(5):e474
17534424
J Biol Chem. 2004 Jun 11;279(24):25430-9
15073168
J Biol Chem. 2001 Apr 13;276(15):12049-54
11116157
Nature. 1990 Dec 6;348(6301):510-4
2174129