title: Pan1 regulates transitions between stages of clathrin-mediated endocytosis.
abstract: Endocytosis is a well-conserved process by which cells invaginate small portions of the plasma membrane to create vesicles containing extracellular and transmembrane cargo proteins. Dozens of proteins and hundreds of specific binding interactions are needed to coordinate and regulate these events. Saccharomyces cerevisiae is a powerful model system with which to study clathrin-mediated endocytosis (CME). Pan1 is believed to be a scaffolding protein due to its interactions with numerous proteins that act throughout the endocytic process. Previous research characterized many Pan1 binding interactions, but due to Pan1's essential nature, the exact mechanisms of Pan1's function in endocytosis have been difficult to define. We created a novel Pan1-degron allele, Pan1-AID, in which Pan1 can be specifically and efficiently degraded in <1 h upon addition of the plant hormone auxin. The loss of Pan1 caused a delay in endocytic progression and weakened connections between the coat/actin machinery and the membrane, leading to arrest in CME. In addition, we determined a critical role for the central region of Pan1 in endocytosis and viability. The regions important for endocytosis and viability can be separated, suggesting that Pan1 may have a distinct role in the cell that is essential for viability.
authors: Department of Biology, Johns Hopkins University, Baltimore, MD 21218.
authors: Mary Katherine
authors: MK
authors: Bradford
authors: Department of Biology, Johns Hopkins University, Baltimore, MD 21218.
authors: Karen
authors: K
authors: Whitworth
authors: Department of Biology, Johns Hopkins University, Baltimore, MD 21218 bwendland@jhu.edu.
authors: Beverly
authors: B
authors: Wendland
mesh_terms: D020816:Amino Acid Motifs; D002966:Clathrin / Q000378:metabolism*; D004705:Endocytosis / Q000502:physiology*; D008840:Microfilament Proteins / Q000378:metabolism* / Q000502:physiology; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000378:metabolism / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism* / Q000502:physiology
chemical_list: D002966:Clathrin; D008840:Microfilament Proteins; C111870:PAN1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins
references: Nat Cell Biol. 2001 Jul;3(7):687-90
references: 11433303
references: J Cell Sci. 2000 Sep;113 ( Pt 18):3309-19
references: 10954428
references: Mol Biol Cell. 2002 Feb;13(2):542-57
references: 11854411
references: Cell. 2003 Nov 14;115(4):475-87
references: 14622601
references: Proc Natl Acad Sci U S A. 2004 Mar 23;101(12):4153-7
references: 15010530
references: Cell Biochem Biophys. 2004;41(2):295-318
references: 15475615
references: J Cell Biol. 1988 May;106(5):1453-61
references: 2836429
references: Cell. 1992 Sep 18;70(6):961-73
references: 1339314
references: Mol Cell Biol. 1995 Dec;15(12):6884-94
references: 8524255
references: EMBO J. 1998 Nov 16;17(22):6541-50
references: 9822599
references: EMBO J. 1999 Mar 1;18(5):1159-71
references: 10064583
references: J Biol Chem. 1999 May 28;274(22):15671-7
references: 10336464
references: EMBO J. 1999 Aug 16;18(16):4383-93
references: 10449404
references: Mol Biol Cell. 1999 Oct;10(10):3521-38
references: 10512884
references: Yeast. 1999 Oct;15(14):1541-53
references: 10514571
references: Nat Cell Biol. 2005 Mar;7(3):246-54
references: 15711538
references: Traffic. 2006 May;7(5):574-88
references: 16643280
references: Mol Biol Cell. 2007 Feb;18(2):658-68
references: 17151356
references: Mol Cell Biol. 2000 Jan;20(1):12-25
references: 10594004
references: Proteins. 2013 Nov;81(11):1944-63
references: 23801378
references: Mol Cell Biol. 2001 Apr;21(7):2449-62
references: 11259593
references: Mol Cell Biol. 1996 Sep;16(9):4897-914
references: 8756649
references: J Biol Chem. 1996 Dec 20;271(51):33123-30
references: 8955161
references: J Cell Biol. 1996 Dec;135(6 Pt 1):1485-500
references: 8978817
references: Mol Cell Biol. 1997 Aug;17(8):4294-304
references: 9234686
references: Genes Dev. 1997 Sep 1;11(17):2239-49
references: 9303539
references: J Cell Biol. 1998 Apr 6;141(1):71-84
references: 9531549
references: Nature. 1998 Jul 9;394(6689):192-5
references: 9671304
references: Yeast. 1998 Jul;14(10):953-61
references: 9717241
references: J Biol Chem. 1998 Nov 20;273(47):31401-7
references: 9813051
references: Cell. 2005 Oct 21;123(2):305-20
references: 16239147
references: Exp Cell Res. 2005 Nov 1;310(2):482-92
references: 16171804
references: Mol Biol Cell. 2007 Aug;18(8):2893-903
references: 17522383
references: FEBS Lett. 2007 Nov 27;581(28):5371-6
references: 17967424
references: Mol Biol Cell. 2007 Dec;18(12):4885-98
references: 17898076
references: Mol Biol Cell. 2008 Jul;19(7):2936-48
references: 18448668
references: Mol Biol Cell. 2009 Nov;20(22):4640-51
references: 19776351
references: Nat Methods. 2009 Dec;6(12):917-22
references: 19915560
references: Traffic. 2010 Sep;11(9):1141-50
references: 20626707
references: J Cell Biol. 2011 Nov 14;195(4):657-71
references: 22065638
references: Mol Biol Cell. 2012 Feb;23(4):687-700
references: 22190739
references: PLoS One. 2012;7(4):e36023
references: 22558309
references: Proc Natl Acad Sci U S A. 2012 Sep 18;109(38):E2533-42
references: 22927393
references: Traffic. 2014 Jan;15(1):43-59
references: 24118836
references: Mol Syst Biol. 2014;10:756
references: 25366307
references: Mol Biol Cell. 2001 Dec;12(12):3759-72
references: 11739778