title: A system for imaging the regulatory noncoding Xist RNA in living mouse embryonic stem cells.
abstract: In mammals, silencing of one of the two X chromosomes in female cells compensates for the different number of X chromosomes between the sexes. The noncoding Xist RNA initiates X chromosome inactivation. Xist spreads from its transcription site over the X chromosome territory and triggers the formation of a repressive chromatin domain. To understand localization of Xist over one X chromosome we aimed to develop a system for investigating Xist in living cells. Here we report successful visualization of transgenically expressed MS2-tagged Xist in mouse embryonic stem cells. Imaging of Xist during an entire cell cycle shows that Xist spreads from a single point to a steady state when the chromosome is covered with a constant amount of Xist. Photobleaching experiments of the established Xist cluster indicate that chromosome-bound Xist is dynamic and turns over on the fully Xist covered chromosome. It appears that in interphase the loss of bound Xist and newly produced Xist are in equilibrium. We also show that the turnover of bound Xist requires transcription, and Xist binding becomes stable when transcription is inhibited. Our data reveal a strategy for visualizing Xist and indicate that spreading over the chromosome might involve dynamic binding and displacement.
authors: Research Institute of Molecular Pathology, 1030 Vienna, Austria.
authors: Karen
authors: K
authors: Ng
authors: Nathalie
authors: N
authors: Daigle
authors: Aurélien
authors: A
authors: Bancaud
authors: Tatsuya
authors: T
authors: Ohhata
authors: Peter
authors: P
authors: Humphreys
authors: Rachael
authors: R
authors: Walker
authors: Jan
authors: J
authors: Ellenberg
authors: Anton
authors: A
authors: Wutz
references: Mol Cell Biol. 2006 Oct;26(19):7167-77
references: 16980619
references: J Cell Sci. 2008 Mar 15;121(Pt 6):887-94
references: 18303053
references: Curr Biol. 2003 Jan 21;13(2):161-167
references: 12546792
references: Nat Genet. 2008 May;40(5):663-9
references: 18425126
references: Prog Mol Subcell Biol. 2004;35:135-50
references: 15113083
references: Mol Cell. 2000 Apr;5(4):695-705
references: 10882105
references: Exp Cell Res. 2010 Mar 10;316(5):679-87
references: 20083102
references: Nature. 1991 May 23;351(6324):329-31
references: 2034279
references: Genes Dev. 1995 Oct 1;9(19):2325-34
references: 7557385
references: Proc Natl Acad Sci U S A. 2002 Jun 25;99(13):8677-82
references: 12072569
references: Cell. 1996 Jul 12;86(1):83-94
references: 8689690
references: J Cell Biol. 1996 Feb;132(3):259-75
references: 8636206
references: Dev Cell. 2010 Sep 14;19(3):469-76
references: 20833368
references: Proc Natl Acad Sci U S A. 2009 Jul 14;106(28):11667-72
references: 19571010
references: Semin Cell Dev Biol. 2003 Dec;14(6):313-8
references: 15015738
references: Proc Natl Acad Sci U S A. 2007 Jun 12;104(24):10104-9
references: 17537922
references: Annu Rev Genet. 2008;42:733-72
references: 18729722
references: Dev Cell. 2005 Jul;9(1):159-65
references: 15992549
references: Gene. 1991 Dec 15;108(2):193-9
references: 1660837
references: Nucleic Acids Res. 1987 Dec 23;15(24):10483-93
references: 3697094
references: J Cell Biol. 2009 Aug 24;186(4):491-507
references: 19704020
references: Genes Dev. 2006 Aug 15;20(16):2223-37
references: 16912274
references: EMBO J. 2006 Jul 12;25(13):3110-22
references: 16763550
references: J Microsc. 2004 Nov;216(Pt 2):131-7
references: 15516224
references: Annu Rev Biophys Biomol Struct. 2007;36:151-69
references: 17477838
references: Proc Natl Acad Sci U S A. 2006 May 16;103(20):7688-93
references: 16682630
references: Epigenetics Chromatin. 2010 May 07;3(1):10
references: 20459652
references: Cytogenet Genome Res. 2002;99(1-4):75-84
references: 12900548
references: PLoS One. 2008 Jun 25;3(6):e2521
references: 18575625
references: Curr Biol. 2006 Aug 8;16(15):1571-8
references: 16890534
references: Science. 1996 Aug 9;273(5276):797-801
references: 8670420
references: Mol Cell. 2006 Mar 3;21(5):617-28
references: 16507360
references: Nature. 1991 Jan 3;349(6304):38-44
references: 1985261
references: Science. 2006 Jun 16;312(5780):1653-5
references: 16778056
references: Cell. 2004 Mar 5;116(5):683-98
references: 15006351
references: Cell. 2007 Jun 29;129(7):1311-23
references: 17604720
references: Curr Opin Cell Biol. 2007 Jun;19(3):311-6
references: 17467967
references: Chromosoma. 2003 Dec;112(4):173-82
references: 14608463
references: Nat Genet. 2002 Feb;30(2):167-74
references: 11780141
references: Nature. 1991 Jan 3;349(6304):82-4
references: 1985270
references: Development. 2004 Mar;131(5):975-82
references: 14973270
references: EMBO J. 2009 Dec 16;28(24):3785-98
references: 19927119
references: Nature. 1991 May 23;351(6324):325-9
references: 2034278
references: Development. 2010 Mar;137(6):935-43
references: 20150277
references: Cell. 1997 Sep 5;90(5):907-16
references: 9298902
references: EMBO J. 2006 Aug 9;25(15):3565-75
references: 16874305
references: Mol Cell. 1998 Oct;2(4):437-45
references: 9809065
references: Nature. 1997 Mar 20;386(6622):272-5
references: 9069284
references: Mol Cell Biol. 1999 Apr;19(4):3156-66
references: 10082582
references: Cell. 2009 Feb 20;136(4):629-41
references: 19239885
references: Genes Dev. 2006 Jul 15;20(14):1848-67
references: 16847345
references: J Biol Chem. 2005 Jan 21;280(3):1720-3
references: 15563465
references: Biochem Cell Biol. 2009 Oct;87(5):759-66
references: 19898525
references: Proc Natl Acad Sci U S A. 2006 May 16;103(20):7706-11
references: 16679409
references: Nature. 2009 Mar 12;458(7235):223-7
references: 19182780
references: Dev Cell. 2009 Apr;16(4):507-16
references: 19386260
references: Science. 2004 Jan 30;303(5658):644-9
references: 14671313