title: Effects of brefeldin A-inhibited guanine nucleotide-exchange (BIG) 1 and KANK1 proteins on cell polarity and directed migration during wound healing.
abstract: Brefeldin A-inhibited guanine nucleotide-exchange protein (BIG) 1 activates class I ADP ribosylation factors (ARFs) by accelerating the replacement of bound GDP with GTP to initiate recruitment of coat proteins for membrane vesicle formation. Among proteins that interact with BIG1, kinesin family member 21A (KIF21A), a plus-end-directed motor protein, moves cargo away from the microtubule-organizing center (MTOC) on microtubules. Because KANK1, a protein containing N-terminal KN, C-terminal ankyrin-repeat, and intervening coiled-coil domains, has multiple actions in cells and also interacts with KIF21A, we explored a possible interaction between it and BIG1. We obtained evidence for a functional and physical association between these proteins, and found that the effects of BIG1 and KANK1 depletion on cell migration in wound-healing assays were remarkably similar. Treatment of cells with BIG1- or KANK1-specific siRNA interfered significantly with directed cell migration and initial orientation of Golgi/MTOC toward the leading edge, which was not mimicked by KIF21A depletion. Although colocalization of overexpressed KANK1 and endogenous BIG1 in HeLa cells was not clear microscopically, their reciprocal immunoprecipitation (IP) is compatible with the presence of small percentages of each protein in the same complexes. Depletion or overexpression of BIG1 protein appeared not to affect KANK1 distribution. Our data identify actions of both BIG1 and KANK1 in regulating cell polarity during directed migration; these actions are consistent with the presence of both BIG1 and KANK1 in dynamic multimolecular complexes that maintain Golgi/MTOC orientation, differ from those that might contain all three proteins (BIG1, KIF21A, and KANK1), and function in directed transport along microtubules.
authors.affiliation: Cardiovascular and Pulmonary Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Chun-Chun
authors.initials: CC
authors.lastname: Li
authors.forename: Jean-Cheng
authors.initials: JC
authors.lastname: Kuo
authors.forename: Clare M
authors.initials: CM
authors.lastname: Waterman
authors.forename: Ryoiti
authors.initials: R
authors.lastname: Kiyama
authors.forename: Joel
authors.initials: J
authors.lastname: Moss
authors.forename: Martha
authors.initials: M
authors.lastname: Vaughan
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D015153:Blotting, Western; D002465:Cell Movement / Q000502:physiology*; D016764:Cell Polarity / Q000502:physiology*; D003598:Cytoskeletal Proteins; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D016547:Kinesins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D025521:Tumor Suppressor Proteins / Q000378:metabolism*; D014945:Wound Healing / Q000502:physiology*
chemical_list: C119087:ARFGEF1 protein, human; D048868:Adaptor Proteins, Signal Transducing; D003598:Cytoskeletal Proteins; D020662:Guanine Nucleotide Exchange Factors; C467739:KANK1 protein, human; C119023:KIF21A protein, human; D034741:RNA, Small Interfering; D025521:Tumor Suppressor Proteins; D016547:Kinesins
references.citation: J Biol Chem. 1999 Apr 30;274(18):12308-15
references.pmid: 10212200
references.citation: Nat Rev Mol Cell Biol. 2009 Jul;10(7):445-57
references.pmid: 19546857
references.citation: Proc Natl Acad Sci U S A. 2000 Mar 14;97(6):2567-72
references.pmid: 10716990
references.citation: J Biol Chem. 2007 Sep 28;282(39):28834-28842
references.pmid: 17640864
references.citation: J Biol Chem. 1999 Jun 18;274(25):17417-23
references.pmid: 10364170
references.citation: Mol Biol Cell. 2005 Oct;16(10):4495-508
references.pmid: 16030262
references.citation: Nature. 2010 Nov 25;468(7323):580-4
references.pmid: 21107430
references.citation: Science. 2006 Sep 15;313(5793):1642-5
references.pmid: 16902090
references.citation: Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2635-40
references.pmid: 16477018
references.citation: J Biol Chem. 2002 Sep 27;277(39):36585-91
references.pmid: 12133830
references.citation: Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):1627-32
references.pmid: 12571360
references.citation: Nat Rev Mol Cell Biol. 2008 Nov;9(11):846-59
references.pmid: 18946474
references.citation: Nat Rev Mol Cell Biol. 2009 Aug;10(8):538-49
references.pmid: 19603038
references.citation: J Cell Biol. 2010 Mar 8;188(5):621-8
references.pmid: 20212314
references.citation: Nat Rev Mol Cell Biol. 2004 Dec;5(12):959-70
references.pmid: 15573134
references.citation: Nat Cell Biol. 2005 Apr;7(4):353-64
references.pmid: 15793564
references.citation: J Cell Sci. 2004 Mar 15;117(Pt 8):1291-300
references.pmid: 15020669
references.citation: J Biol Chem. 1998 Aug 21;273(34):21431-4
references.pmid: 9705267
references.citation: Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3357-61
references.pmid: 18292223
references.citation: Mol Biol Cell. 2002 Jan;13(1):119-33
references.pmid: 11809827
references.citation: J Cell Sci. 2009 Sep 15;122(Pt 18):3215-23
references.pmid: 19726631
references.citation: Proc Natl Acad Sci U S A. 2004 Mar 2;101(9):2752-7
references.pmid: 14973189
references.citation: Proc Natl Acad Sci U S A. 2010 Jul 6;107(27):12145-50
references.pmid: 20566876
references.citation: Proc Natl Acad Sci U S A. 2007 Jan 23;104(4):1230-5
references.pmid: 17227842
references.citation: Cell. 2005 May 6;121(3):451-63
references.pmid: 15882626
references.citation: Proc Natl Acad Sci U S A. 1999 Jul 6;96(14):7968-73
references.pmid: 10393931
references.citation: Biochem Biophys Res Commun. 2005 May 20;330(4):1247-53
references.pmid: 15823577
references.citation: J Cell Biol. 2009 Jan 26;184(2):253-67
references.pmid: 19171758
references.citation: Mol Biol Cell. 2008 Jun;19(6):2650-60
references.pmid: 18417613
references.citation: J Cell Sci. 2006 Oct 1;119(Pt 19):4002-10
references.pmid: 16968744
references.citation: Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):12856-60
references.pmid: 8917509
references.citation: Dev Biol. 2004 Jan 1;265(1):23-32
references.pmid: 14697350
references.citation: Curr Opin Cell Biol. 2003 Feb;15(1):67-72
references.pmid: 12517706
references.citation: Nat Rev Mol Cell Biol. 2008 Apr;9(4):273-84
references.pmid: 18354421
references.citation: Nat Cell Biol. 2006 Dec;8(12):1383-8
references.pmid: 17086175
references.citation: Science. 2003 Dec 5;302(5651):1704-9
references.pmid: 14657486
references.citation: J Biol Chem. 2005 Mar 18;280(11):10128-34
references.pmid: 15644318
references.citation: Proc Natl Acad Sci U S A. 1982 Apr;79(8):2603-7
references.pmid: 7045867
references.citation: Nat Methods. 2008 Feb;5(2):155-7
references.pmid: 18193054
references.citation: Cell Mol Life Sci. 2009 Aug;66(16):2651-9
references.pmid: 19554261
references.citation: Nat Cell Biol. 2005 May;7(5):483-92
references.pmid: 15821732
references.citation: Proc Natl Acad Sci U S A. 2008 Dec 2;105(48):18788-93
references.pmid: 19020088
references.citation: Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2683-8
references.pmid: 16467138
references.citation: PLoS One. 2010 Mar 25;5(3):e9898
references.pmid: 20360857
references.citation: Mol Biol Cell. 2004 Dec;15(12):5283-94
references.pmid: 15385626
references.citation: J Cell Biol. 2008 May 5;181(3):537-49
references.pmid: 18458160
references.citation: Proc Natl Acad Sci U S A. 2010 Jul 6;107(27):12399-404
references.pmid: 20562345
references.citation: Biochem Biophys Res Commun. 2009 Sep 4;386(4):639-44
references.pmid: 19559006
references.citation: Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3201-6
references.pmid: 17360629
references.citation: Trends Cell Biol. 2000 Feb;10(2):60-7
references.pmid: 10652516
references.citation: Trends Cell Biol. 2010 Jun;20(6):319-28
references.pmid: 20399659
references.citation: Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2322-7
references.pmid: 12606707
references.citation: Proc Natl Acad Sci U S A. 2005 Feb 22;102(8):2784-9
references.pmid: 15705715
references.citation: IUBMB Life. 2009 Apr;61(4):394-406
references.pmid: 19319965