Yeast Golgi-localized, gamma-Ear-containing, ADP-ribosylation factor-binding proteins are but adaptor protein-1 is not required for cell-free transport of membrane proteins from the trans-Golgi network to the prevacuolar compartment.
Golgi-localized, gamma-Ear-containing, ADP-ribosylation factor-binding proteins (GGAs) and adaptor protein-1 (AP-1) mediate clathrin-dependent trafficking of transmembrane proteins between the trans-Golgi network (TGN) and endosomes. In yeast, the vacuolar sorting receptor Vps10p follows a direct pathway from the TGN to the late endosome/prevacuolar compartment (PVC), whereas, the processing protease Kex2p partitions between the direct pathway and an indirect pathway through the early endosome. To examine the roles of the Ggas and AP-1 in TGN-PVC transport, we used a cell-free assay that measures delivery to the PVC of either Kex2p or a chimeric protein (K-V), in which the Vps10p cytosolic tail replaces the Kex2p tail. Either antibody inhibition or dominant-negative Gga2p completely blocked K-V transport but only partially blocked Kex2p transport. Deletion of APL2, encoding the beta subunit of AP-1, did not affect K-V transport but partially blocked Kex2p transport. Residual Kex2p transport seen with apl2Delta membranes was insensitive to dominant-negative Gga2p, suggesting that the apl2Delta mutation causes Kex2p to localize to a compartment that precludes Gga-dependent trafficking. These results suggest that yeast Ggas facilitate the specific and direct delivery of Vps10p and Kex2p from the TGN to the PVC and that AP-1 modulates Kex2p trafficking through a distinct pathway, presumably involving the early endosome.
Abazeed
Mohamed E
ME
Department of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109, USA.
Fuller
Robert S
RS
D020727:ADP-Ribosylation Factors / Q000302:isolation & purification / Q000378:metabolism*; D033961:Adaptor Protein Complex 1 / Q000378:metabolism*; D033942:Adaptor Proteins, Vesicular Transport / Q000302:isolation & purification / Q000378:metabolism*; D017353:Gene Deletion; D005799:Genes, Dominant; D008565:Membrane Proteins / Q000378:metabolism*; D021381:Protein Transport; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism*; D021601:trans-Golgi Network / Q000378:metabolism*
D033961:Adaptor Protein Complex 1; D033942:Adaptor Proteins, Vesicular Transport; C407306:GGA adaptor proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D020727:ADP-Ribosylation Factors
Mol Cell Biol. 1996 Nov;16(11):6208-17
8887651
J Cell Biol. 2001 Dec 10;155(6):969-78
11739408
Yeast. 1998 Jul;14(10):953-61
9717241
Mol Biol Cell. 1992 Dec;3(12):1353-71
1493334
FEBS Lett. 2002 Jan 30;511(1-3):155-8
11821067
J Cell Biol. 2000 Oct 30;151(3):587-600
11062260
Mol Biol Cell. 1995 Sep;6(9):1089-102
8534908
J Cell Sci. 1995 Apr;108 ( Pt 4):1605-15
7615679
Mol Biol Cell. 2004 Jul;15(7):3196-209
15090613
Annu Rev Cell Dev Biol. 2004;20:153-91
15473838
J Cell Sci. 2003 Mar 1;116(Pt 5):763-71
12571274
Nature. 2002 Feb 21;415(6874):933-7
11859375
Mol Biol Cell. 2001 Jun;12(6):1885-96
11408593
Anal Biochem. 1991 Feb 1;192(2):262-7
1852137
Nucleic Acids Res. 2002 Mar 15;30(6):e23
11884642
Nat Rev Mol Cell Biol. 2004 Jan;5(1):23-32
14708007
J Cell Biol. 2000 Apr 3;149(1):81-94
10747089
J Cell Biol. 2006 May 22;173(4):615-26
16702232
J Cell Biol. 1996 May;133(3):529-41
8636229
Mol Biol Cell. 2001 Feb;12(2):475-85
11179429
Gene. 1995 Apr 14;156(1):119-22
7737504
Dev Cell. 2002 Mar;2(3):283-94
11879634
Science. 2002 Sep 6;297(5587):1700-3
12215646
Yeast. 1994 Apr;10(4):441-9
7941730
Mol Biol Cell. 2008 May;19(5):1991-2002
18287542
Traffic. 2000 Mar;1(3):259-69
11208109
Nat Struct Biol. 2003 May;10(5):386-93
12679809
Mol Biol Cell. 1999 Nov;10(11):3643-59
10564262
Science. 2001 Jun 1;292(5522):1716-8
11387476
Mol Biol Cell. 2007 Jul;18(7):2646-55
17494868
Mol Biol Cell. 2003 Apr;14(4):1319-33
12686590
Mol Cell Biol. 1990 Dec;10(12):6690-9
2123295
Cell. 2004 Jan 23;116(2):153-66
14744428
Proc Natl Acad Sci U S A. 1986 May;83(10):3248-52
3517855
J Cell Biol. 1997 Oct 6;139(1):23-36
9314526
J Biol Chem. 2001 Jul 27;276(30):28541-5
11390366
EMBO J. 1994 Apr 1;13(7):1706-17
8157009
Bioessays. 2001 Apr;23(4):333-43
11268039
J Cell Biol. 2000 Apr 3;149(1):67-80
10747088
Genetics. 1989 May;122(1):19-27
2659436
Science. 2001 Jun 1;292(5522):1712-6
11387475
J Biol Chem. 2005 Feb 11;280(6):4442-50
15572353
J Biol Chem. 2004 Nov 19;279(47):48767-73
15364946
J Cell Biol. 1991 May;113(3):527-38
2016334
Mol Biol Cell. 1995 Jan;6(1):41-58
7749194
Mol Biol Cell. 1996 Apr;7(4):579-94
8730101
Mol Biol Cell. 2001 Nov;12(11):3573-88
11694590