title: Activation of Lyn tyrosine kinase through decreased membrane cholesterol levels during a change in its membrane distribution upon cell detachment.
abstract: Cellular membranes, which can serve as scaffolds for signal transduction, dynamically change their characteristics upon cell detachment. Src family kinases undergo post-translational lipid modification and are involved in a wide range of signaling events at the plasma membrane, such as cell proliferation, cell adhesion, and survival. Previously, we showed the differential membrane distributions among the members of Src family kinases by sucrose density gradient fractionation. However, little is known about the regulation of the membrane distribution of Src family kinases upon cell detachment. Here, we show that cell detachment shifts the main peak of the membrane distribution of Lyn, a member of Src family kinase, from the low density to the high density membrane fractions and enhances the kinase activity of Lyn. The change in Lyn distribution upon cell detachment involves both dynamin activity and a decrease in membrane cholesterol. Cell detachment activates Lyn through decreased membrane cholesterol levels during a change in its membrane distribution. Furthermore, cholesterol incorporation decreases Lyn activity and reduces the viability of suspension cells. These results suggest that cell detachment-induced Lyn activation through the change in the membrane distribution of Lyn plays an important role in survival of suspension cells.
authors.affiliation: Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Morinaga
authors.affiliation: Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Abe
authors.affiliation: Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Nakayama
authors.affiliation: Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan.
authors.forename: Noritaka
authors.initials: N
authors.lastname: Yamaguchi
authors.affiliation: Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan. Electronic address: nyama@faculty.chiba-u.jp.
authors.forename: Naoto
authors.initials: N
authors.lastname: Yamaguchi
mesh_terms: D002448:Cell Adhesion; D002462:Cell Membrane / Q000201:enzymology*; D002470:Cell Survival; D002784:Cholesterol / Q000494:pharmacology / Q000502:physiology*; D034281:Dynamins / Q000037:antagonists & inhibitors / Q000378:metabolism; D004789:Enzyme Activation; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D006835:Hydrazones / Q000494:pharmacology; D019308:Palmitic Acid / Q000378:metabolism; D021381:Protein Transport; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: D006835:Hydrazones; C511794:N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide; D019308:Palmitic Acid; D002784:Cholesterol; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases; D034281:Dynamins
keywords: Cell Detachment; Cholesterol; Kinase Activity; Lyn; Membrane Function; Membrane Segment; Src; Subcellular Fractionation; Tyrosine-Protein Kinase (Tyrosine Kinase)
references.citation: QJM. 2012 Apr;105(4):383-8
references.pmid: 22156736
references.citation: Exp Cell Res. 2007 Jul 15;313(12):2651-66
references.pmid: 17537435
references.citation: Semin Cell Dev Biol. 2012 Apr;23(2):126-44
references.pmid: 22309841
references.citation: J Pathol. 2010 Apr;220(5):574-85
references.pmid: 20146241
references.citation: J Cell Biol. 2004 Jun 7;165(5):641-52
references.pmid: 15173188
references.citation: Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10104-8
references.pmid: 7479734
references.citation: Mol Biol Cell. 2011 Sep;22(18):3456-64
references.pmid: 21795400
references.citation: Mol Carcinog. 2010 Dec;49(12):1007-21
references.pmid: 20945416
references.citation: J Biol Chem. 1995 May 19;270(20):12170-6
references.pmid: 7744867
references.citation: Mol Cell Biol. 2009 Dec;29(24):6462-72
references.pmid: 19822664
references.citation: J Biol Chem. 2004 Feb 13;279(7):5975-83
references.pmid: 14613929
references.citation: Nat Rev Mol Cell Biol. 2007 Aug;8(8):633-44
references.pmid: 17643125
references.citation: J Biol Chem. 2012 Jul 20;287(30):24905-15
references.pmid: 22689581
references.citation: Science. 2014 Mar 28;343(6178):1445-6
references.pmid: 24675946
references.citation: J Biol Chem. 2003 Jun 6;278(23):20746-52
references.pmid: 12670955
references.citation: J Cell Sci. 2002 Mar 15;115(Pt 6):1331-40
references.pmid: 11884532
references.citation: J Biol Chem. 2004 Mar 26;279(13):13110-8
references.pmid: 14711821
references.citation: J Clin Invest. 1975 Jan;55(1):115-26
references.pmid: 162782
references.citation: Science. 2004 Feb 6;303(5659):839-42
references.pmid: 14764880
references.citation: Mol Cell. 2004 Feb 13;13(3):341-55
references.pmid: 14967142
references.citation: Exp Cell Res. 2006 Apr 15;312(7):1205-17
references.pmid: 16527271
references.citation: Oncogene. 2005 Mar 3;24(10):1727-37
references.pmid: 15674335
references.citation: J Cell Sci. 2013 Jan 1;126(Pt 1):21-9
references.pmid: 23516327
references.citation: J Biol Chem. 2014 May 2;289(18):12313-29
references.pmid: 24634213
references.citation: Cell. 2011 Oct 14;147(2):275-92
references.pmid: 22000009
references.citation: J Cell Biol. 2000 Apr 17;149(2):431-46
references.pmid: 10769034
references.citation: J Chromatogr B Biomed Sci Appl. 1999 Feb 5;722(1-2):89-102
references.pmid: 10068135
references.citation: J Cell Biol. 1968 May;37(2):482-513
references.pmid: 4297786
references.citation: Cytotechnology. 2010 Jan;62(1):73-82
references.pmid: 20309632
references.citation: J Cell Biol. 1969 Oct;43(1):59-79
references.pmid: 4241907
references.citation: Cell Motil Cytoskeleton. 2007 Mar;64(3):199-216
references.pmid: 17238130
references.citation: Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3165-70
references.pmid: 17360623
references.citation: Mol Cell. 2008 May 23;30(4):426-36
references.pmid: 18498747
references.citation: Nature. 1978 Jun 1;273(5661):345-9
references.pmid: 661946
references.citation: Methods Enzymol. 2003;370:430-44
references.pmid: 14712665
references.citation: Biochim Biophys Acta. 2009 Oct;1790(10):1345-52
references.pmid: 19619611
references.citation: Biochim Biophys Acta. 1996 Jun 7;1287(2-3):121-49
references.pmid: 8672527
references.citation: J Cell Biol. 2008 Jul 28;182(2):367-79
references.pmid: 18644892
references.citation: Oncogene. 2004 Dec 2;23(56):9052-61
references.pmid: 15489898
references.citation: J Exp Med. 1956 Feb 1;103(2):273-83
references.pmid: 13286432
references.citation: Cancer Metastasis Rev. 2003 Dec;22(4):337-58
references.pmid: 12884910
references.citation: Genes Dev. 2012 Jan 1;26(1):54-68
references.pmid: 22215811
references.citation: Biochemistry. 1995 Oct 24;34(42):13784-93
references.pmid: 7577971
references.citation: Science. 2010 Jan 1;327(5961):46-50
references.pmid: 20044567
references.citation: Cancer Res. 2004 Feb 1;64(3):1058-66
references.pmid: 14871838
references.citation: Front Biosci. 1997 Oct 15;2:d501-18
references.pmid: 9331427
references.citation: J Cell Sci. 2011 Aug 15;124(Pt 16):2763-76
references.pmid: 21807941
references.citation: Exp Cell Res. 2013 Jun 10;319(10):1382-97
references.pmid: 23562843
references.citation: Proc Natl Acad Sci U S A. 2012 Mar 27;109(13):4869-74
references.pmid: 22411821
references.citation: Cancer Res. 2010 Mar 15;70(6):2296-306
references.pmid: 20215510
references.citation: J Cell Sci. 2009 Apr 1;122(Pt 7):965-75
references.pmid: 19258394
references.citation: Curr Opin Cell Biol. 2001 Oct;13(5):555-62
references.pmid: 11544023