title: Phosphorylation of conserved PIN motifs directs Arabidopsis PIN1 polarity and auxin transport.
abstract: Polar cell-to-cell transport of auxin by plasma membrane-localized PIN-FORMED (PIN) auxin efflux carriers generates auxin gradients that provide positional information for various plant developmental processes. The apical-basal polar localization of the PIN proteins that determines the direction of auxin flow is controlled by reversible phosphorylation of the PIN hydrophilic loop (PINHL). Here, we identified three evolutionarily conserved TPRXS(N/S) motifs within the PIN1HL and proved that the central Ser residues were phosphorylated by the PINOID (PID) kinase. Loss-of-phosphorylation PIN1:green fluorescent protein (GFP) (Ser to Ala) induced inflorescence defects, correlating with their basal localization in the shoot apex, and induced internalization of PIN1:GFP during embryogenesis, leading to strong embryo defects. Conversely, phosphomimic PIN1:GFP (Ser to Glu) showed apical localization in the shoot apex but did not rescue pin1 inflorescence defects. Both loss-of-phosphorylation and phosphomimic PIN1:GFP proteins were insensitive to PID overexpression. The basal localization of loss-of-phosphorylation PIN1:GFP increased auxin accumulation in the root tips, partially rescuing PID overexpression-induced root collapse. Collectively, our data indicate that reversible phosphorylation of the conserved Ser residues in the PIN1HL by PID (and possibly by other AGC kinases) is required and sufficient for proper PIN1 localization and is thus essential for generating the differential auxin distribution that directs plant development.
authors.affiliation: Department of Molecular and Developmental Genetics, Institute of Biology, Leiden University, 2333 EB Leiden, The Netherlands.
authors.forename: Fang
authors.initials: F
authors.lastname: Huang
authors.forename: Marcelo Kemel
authors.initials: MK
authors.lastname: Zago
authors.forename: Lindy
authors.initials: L
authors.lastname: Abas
authors.forename: Arnoud
authors.initials: A
authors.lastname: van Marion
authors.forename: Carlos Samuel
authors.initials: CS
authors.lastname: Galván-Ampudia
authors.forename: Remko
authors.initials: R
authors.lastname: Offringa
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D018744:DNA, Plant / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D007210:Indoleacetic Acids / Q000378:metabolism*; D056626:Inflorescence / Q000254:growth & development; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D010937:Plant Growth Regulators / Q000378:metabolism*; D054730:Protein Interaction Domains and Motifs; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: D029681:Arabidopsis Proteins; D018744:DNA, Plant; D007210:Indoleacetic Acids; D026901:Membrane Transport Proteins; C116363:PIN1 protein, Arabidopsis; D010937:Plant Growth Regulators; C405165:PINOID protein, Arabidopsis; D017346:Protein Serine-Threonine Kinases
references.citation: Plant Physiol. 1998 Feb 1;116(2):833-44
references.pmid: 9490775
references.citation: Nature. 2005 Jun 30;435(7046):1251-6
references.pmid: 15988527
references.citation: Plant Cell Physiol. 2005 Oct;46(10):1674-81
references.pmid: 16085936
references.citation: Nat Cell Biol. 2006 Mar;8(3):249-56
references.pmid: 16489343
references.citation: Development. 2004 Oct;131(20):5021-30
references.pmid: 15371311
references.citation: Cell. 2007 Sep 21;130(6):1044-56
references.pmid: 17889649
references.citation: Science. 1998 Dec 18;282(5397):2226-30
references.pmid: 9856939
references.citation: Science. 1990 May 11;248(4956):742-5
references.pmid: 2110383
references.citation: Plant J. 2008 Jan;53(2):380-92
references.pmid: 18047472
references.citation: Anal Biochem. 1991 Feb 1;192(2):262-7
references.pmid: 1852137
references.citation: Curr Biol. 2008 Apr 8;18(7):526-31
references.pmid: 18394892
references.citation: Cell. 2000 Feb 18;100(4):469-78
references.pmid: 10693763
references.citation: Development. 2005 Oct;132(20):4521-31
references.pmid: 16192309
references.citation: Plant J. 1998 Dec;16(6):735-43
references.pmid: 10069079
references.citation: Science. 2006 May 12;312(5775):883
references.pmid: 16601151
references.citation: Endocr Rev. 2001 Apr;22(2):153-83
references.pmid: 11294822
references.citation: Plant J. 2006 Mar;45(5):752-64
references.pmid: 16460509
references.citation: Plant Cell. 2004 Sep;16(9):2394-405
references.pmid: 15308754
references.citation: Plant Cell. 2001 Jul;13(7):1683-97
references.pmid: 11449059
references.citation: Plant Physiol. 2009 Jun;150(2):722-35
references.pmid: 19363095
references.citation: Mol Cell Proteomics. 2007 Jul;6(7):1198-214
references.pmid: 17317660
references.citation: Curr Biol. 2005 Nov 8;15(21):1899-911
references.pmid: 16271866
references.citation: Nature. 2003 Nov 20;426(6964):255-60
references.pmid: 14628043
references.citation: Plant Cell Physiol. 2007 Mar;48(3):540-9
references.pmid: 17303594
references.citation: Cell. 1999 Nov 24;99(5):463-72
references.pmid: 10589675
references.citation: Development. 2009 Feb;136(4):627-36
references.pmid: 19168677
references.citation: Plant Cell. 2004 Dec;16(12):3386-99
references.pmid: 15539472
references.citation: J Mol Biol. 1999 Dec 17;294(5):1351-62
references.pmid: 10600390
references.citation: Cell. 2003 Nov 26;115(5):591-602
references.pmid: 14651850
references.citation: Plant Physiol. 2007 Jun;144(2):1000-11
references.pmid: 17449648
references.citation: FEBS Lett. 2006 Feb 13;580(4):1094-102
references.pmid: 16359667
references.citation: Anal Biochem. 2010 Jun 15;401(2):217-27
references.pmid: 20193653
references.citation: Nature. 2008 Dec 18;456(7224):962-6
references.pmid: 18953331
references.citation: Nature. 2009 May 28;459(7246):583-6
references.pmid: 19478783
references.citation: Genes Dev. 2006 Oct 15;20(20):2902-11
references.pmid: 17043314
references.citation: EMBO J. 1998 Dec 1;17(23):6903-11
references.pmid: 9843496
references.citation: Development. 2001 Oct;128(20):4057-67
references.pmid: 11641228
references.citation: Proc Natl Acad Sci U S A. 2010 Jan 12;107(2):918-22
references.pmid: 20080776
references.citation: Proc Natl Acad Sci U S A. 2001 Jun 5;98(12):6969-74
references.pmid: 11371609
references.citation: Plant Cell. 1991 Jul;3(7):677-684
references.pmid: 12324609
references.citation: Science. 1999 Oct 8;286(5438):316-8
references.pmid: 10514379
references.citation: Nature. 2009 Jun 25;459(7250):1136-40
references.pmid: 19506555
references.citation: Plant Cell. 2009 Dec;21(12):3839-49
references.pmid: 20040538
references.citation: Plant Cell Physiol. 2009 Mar;50(3):652-7
references.pmid: 19153156
references.citation: Trends Plant Sci. 2007 Dec;12(12):541-7
references.pmid: 18024140
references.citation: Trends Plant Sci. 2003 Sep;8(9):424-31
references.pmid: 13678909
references.citation: Nature. 2001 Sep 27;413(6854):425-8
references.pmid: 11574889
references.citation: Cell. 2002 Mar 8;108(5):661-73
references.pmid: 11893337
references.citation: Plant Physiol. 2006 Sep;142(1):254-64
references.pmid: 16844839
references.citation: Nature. 2002 Feb 14;415(6873):806-9
references.pmid: 11845211
references.citation: Trends Plant Sci. 2005 Apr;10(4):170-7
references.pmid: 15817418
references.citation: Nature. 2003 Nov 13;426(6963):147-53
references.pmid: 14614497
references.citation: EMBO J. 1996 May 1;15(9):2115-24
references.pmid: 8641277
references.citation: Nature. 2005 Jan 6;433(7021):39-44
references.pmid: 15635403
references.citation: Cell. 2003 Jan 24;112(2):219-30
references.pmid: 12553910
references.citation: Science. 2004 Oct 29;306(5697):862-5
references.pmid: 15514156
references.citation: Plant Physiol. 2003 Jul;132(3):1623-30
references.pmid: 12857841
references.citation: Proc Natl Acad Sci U S A. 2008 Dec 30;105(52):21017-22
references.pmid: 19075219