LGN regulates mitotic spindle orientation during epithelial morphogenesis.
Coordinated cell polarization and mitotic spindle orientation are thought to be important for epithelial morphogenesis. Whether spindle orientation is indeed linked to epithelial morphogenesis and how it is controlled at the molecular level is still unknown. Here, we show that the NuMA- and Galpha-binding protein LGN is required for directing spindle orientation during cystogenesis of MDCK cells. LGN localizes to the lateral cell cortex, and is excluded from the apical cell cortex of dividing cells. Depleting LGN, preventing its cortical localization, or disrupting its interaction with endogenous NuMA or Galpha proteins all lead to spindle misorientation and abnormal cystogenesis. Moreover, artificial mistargeting of endogenous LGN to the apical membrane results in a near 90 degrees rotation of the spindle axis and profound cystogenesis defects that are dependent on cell division. The normal apical exclusion of LGN during mitosis appears to be mediated by atypical PKC. Thus, cell polarization-mediated spatial restriction of spindle orientation determinants is critical for epithelial morphogenesis.
Zheng
Zhen
Z
Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, GA 30912, USA.
Zhu
Huabin
H
Wan
Qingwen
Q
Liu
Jing
J
Xiao
Zhuoni
Z
Siderovski
David P
DP
Du
Quansheng
Q
D000818:Animals; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D016764:Cell Polarity; D049109:Cell Proliferation; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology; D004848:Epithelium / Q000196:embryology*; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D009024:Morphogenesis / Q000502:physiology*; D034521:Nuclear Matrix-Associated Proteins / Q000235:genetics / Q000378:metabolism; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism* / Q000648:ultrastructure
D044385:GTP-Binding Protein alpha Subunits; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D034521:Nuclear Matrix-Associated Proteins; C115015:PKC-3 protein; D011493:Protein Kinase C
Biochim Biophys Acta. 2005 Sep 10;1745(2):254-64
15946753
Science. 2003 Jun 20;300(5627):1957-61
12750478
Nat Cell Biol. 2009 Apr;11(4):365-74
19337318
J Cell Biol. 2007 Oct 8;179(1):15-22
17908918
Mol Biol Cell. 2003 Feb;14(2):748-63
12589067
Nat Cell Biol. 2008 Jan;10(1):93-101
18084280
Nat Neurosci. 2007 Nov;10(11):1440-8
17934458
Curr Opin Cell Biol. 2003 Feb;15(1):73-81
12517707
Curr Biol. 2002 Nov 19;12(22):1928-33
12445386
Curr Biol. 2005 Sep 20;15(18):1640-4
16169485
Dev Cell. 2006 Jun;10(6):731-42
16740476
J Cell Sci. 2004 Dec 15;117(Pt 26):6391-400
15561764
Dev Cell. 2005 Apr;8(4):461-5
15809029
Science. 1999 Oct 29;286(5441):971-4
10542155
Curr Opin Cell Biol. 2006 Dec;18(6):648-57
17046223
J Cell Biol. 2008 Nov 17;183(4):625-33
19001128
Cell. 1996 Nov 1;87(3):447-58
8898198
Nat Rev Mol Cell Biol. 2008 Nov;9(11):887-901
18946477
J Cell Biol. 1997 Sep 8;138(5):1055-66
9281583
Proc Natl Acad Sci U S A. 2005 Nov 15;102(46):16584-9
16275912
Int Rev Cytol. 1971;31:169-213
4400359
Nat Rev Mol Cell Biol. 2002 Jul;3(7):531-7
12094219
Cell. 2007 Jan 26;128(2):383-97
17254974
J Cell Biol. 2002 Apr 1;157(1):161-72
11927608
Methods. 2003 Jul;30(3):256-68
12798140
Mol Biol Cell. 2003 Aug;14(8):3144-55
12925752
Nat Cell Biol. 2001 Dec;3(12):1069-75
11781568
Annu Rev Biochem. 2004;73:925-51
15189163
J Biol Chem. 2001 Jan 12;276(2):1585-93
11042168
Cell. 2009 Sep 18;138(6):1150-63
19766567
J Biol Chem. 2005 Feb 25;280(8):6761-5
15632202
Mol Cell Biol. 2009 May;29(10):2816-27
19273597
Dev Cell. 2007 Dec;13(6):796-811
18061563
Science. 2003 Jul 25;301(5632):518-21
12881570
Semin Cell Dev Biol. 2009 Oct;20(8):972-7
19447051
Nat Cell Biol. 2006 Jun;8(6):586-93
16648846
Cell. 2004 Nov 12;119(4):503-16
15537540
J Cell Sci. 2009 May 15;122(Pt 10):1595-606
19401335
Cell. 2005 Jul 15;122(1):119-31
16009138
Nature. 2007 May 24;447(7143):493-6
17495931
Development. 2006 Feb;133(3):529-36
16396904
Nat Rev Cancer. 2005 Sep;5(9):675-88
16148884
Nat Cell Biol. 2005 Dec;7(12):1179-90
16299499
J Cell Biol. 2001 Feb 5;152(3):425-34
11157972
Curr Biol. 2004 Aug 24;14(16):R674-85
15324689
EMBO J. 2007 Mar 21;26(6):1487-98
17318179
Curr Opin Cell Biol. 2004 Apr;16(2):195-205
15196564
J Cell Sci. 2003 Jul 15;116(Pt 14):2895-906
12771187
Nat Genet. 2008 Aug;40(8):1010-5
18604206
Cell. 2005 Dec 29;123(7):1323-35
16377571
Proc Natl Acad Sci U S A. 2007 Sep 4;104(36):14306-11
17726110
Dev Biol. 2008 Mar 1;315(1):42-54
18234174
Mol Biol Cell. 2009 Aug;20(16):3740-50
19553471
Nat Cell Biol. 2006 Jun;8(6):594-600
16648843
J Biol Chem. 2008 Dec 26;283(52):36698-710
18984596