title: An ATPase-deficient variant of the SNF2 family member HELLS shows altered dynamics at pericentromeric heterochromatin.
abstract: The HELLS (helicase, lymphoid specific, also known as lymphoid-specific helicase) protein is related to the SNF2 (sucrose non-fermentable 2) family of chromatin remodeling ATPases. It is required for efficient DNA methylation in mammals, particularly at heterochromatin-located repetitive sequences. In this study, we investigated the interaction of HELLS with chromatin and used an ATPase-deficient HELLS variant to address the role of ATP hydrolysis in this process. Chromatin fractionation experiments demonstrated that, in the absence of the ATPase activity, HELLS is retained at the nuclear matrix compartment, defined in part by lamin B1. Microscopy studies revealed a stronger association of the ATPase-deficient mutant with heterochromatin. These results were further supported by fluorescence recovery after photobleaching measurements, which showed that, at heterochromatic sites, wild-type HELLS is very dynamic, with a recovery half-time of 0.8s and a mobile protein fraction of 61%. In contrast, the ATPase-deficient mutant displayed 4.5-s recovery half-time and a reduction in the mobile fraction to 30%. We also present evidence suggesting that, in addition to the ATPase activity, a functional H3K9me3 signaling pathway contributes to an efficient release of HELLS from pericentromeric chromatin. Overall, our results show that a functional ATPase activity is not required for the recruitment of HELLS to heterochromatin, but it is important for the release of the enzyme from these sites.
authors.affiliation: Institute of Biochemistry, Stuttgart University, Pfaffenwaldring 55, D-70569 Stuttgart, Germany.
authors.forename: Cristiana
authors.initials: C
authors.lastname: Lungu
authors.affiliation: Mouse Cancer Genetics Program, Basic Science Program, Leidos Biomedical Research, Inc., National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Muegge
authors.affiliation: Institute of Biochemistry, Stuttgart University, Pfaffenwaldring 55, D-70569 Stuttgart, Germany.
authors.forename: Albert
authors.initials: A
authors.lastname: Jeltsch
authors.affiliation: Institute of Biochemistry, Stuttgart University, Pfaffenwaldring 55, D-70569 Stuttgart, Germany. Electronic address: renata.jurkowska@ibc.uni-stuttgart.de.
authors.forename: Renata Z
authors.initials: RZ
authors.lastname: Jurkowska
mesh_terms: D000251:Adenosine Triphosphatases / Q000172:deficiency*; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D036681:Fluorescence Recovery After Photobleaching; D006570:Heterochromatin / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D008780:Methyltransferases / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D012097:Repressor Proteins / Q000502:physiology*; D014158:Transcription, Genetic
chemical_list: D006570:Heterochromatin; D006657:Histones; D050505:Mutant Proteins; D012097:Repressor Proteins; C498700:Suv39h1 protein, mouse; D008780:Methyltransferases; D000251:Adenosine Triphosphatases; D004265:DNA Helicases; C487055:Hells protein, mouse
keywords: ATP hydrolysis; chromatin remodeling; fluorescence recovery after photobleaching; heterochromatin; protein dynamics
references.citation: Genome Res. 2011 Jan;21(1):83-94
references.pmid: 21149390
references.citation: Nat Rev Genet. 2012 Apr 03;13(5):343-57
references.pmid: 22473383
references.citation: Genome Res. 2014 Oct;24(10):1613-23
references.pmid: 25170028
references.citation: Nucleic Acids Res. 1996 Oct 1;24(19):3685-92
references.pmid: 8871545
references.citation: Mol Cell Biol. 2008 Jan;28(1):215-26
references.pmid: 17967891
references.citation: Cell. 2013 Mar 28;153(1):193-205
references.pmid: 23540698
references.citation: Bioessays. 2003 Dec;25(12):1192-200
references.pmid: 14635254
references.citation: Genomics. 1998 Dec 15;54(3):477-83
references.pmid: 9878251
references.citation: Mol Cell Biol. 2003 Dec;23(23):8416-28
references.pmid: 14612388
references.citation: Trends Biochem Sci. 2014 Jul;39(7):310-8
references.pmid: 24947342
references.citation: Curr Opin Struct Biol. 2012 Apr;22(2):225-33
references.pmid: 22445226
references.citation: Proc Natl Acad Sci U S A. 2011 Apr 5;108(14):5626-31
references.pmid: 21427231
references.citation: Biochim Biophys Acta. 2014 Aug;1839(8):728-36
references.pmid: 24583555
references.citation: J Cell Biol. 2011 Mar 7;192(5):735-50
references.pmid: 21357745
references.citation: Gene. 1996 Mar 9;169(2):203-7
references.pmid: 8647447
references.citation: Nature. 2001 May 10;411(6834):212-4
references.pmid: 11346800
references.citation: Curr Biol. 2003 Jul 15;13(14):1192-200
references.pmid: 12867029
references.citation: PLoS One. 2010 Feb 11;5(2):e9163
references.pmid: 20161795
references.citation: Cancer Res. 2000 Jul 1;60(13):3612-22
references.pmid: 10910076
references.citation: Genome Biol. 2013 Dec 24;14(12):R146
references.pmid: 24367978
references.citation: Nucleic Acids Res. 2004 Sep 24;32(17):5019-28
references.pmid: 15448183
references.citation: Cell. 1997 Apr 18;89(2):195-204
references.pmid: 9108475
references.citation: Genes Dev. 2001 Nov 15;15(22):2940-4
references.pmid: 11711429
references.citation: Nature. 2008 Jun 12;453(7197):948-51
references.pmid: 18463634
references.citation: Nat Cell Biol. 2006 Dec;8(12):1448-54
references.pmid: 17115026
references.citation: Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4772-7
references.pmid: 10781083
references.citation: Biochim Biophys Acta. 2012 Jul;1819(7):757-62
references.pmid: 22381140
references.citation: Nat Genet. 2002 Mar;30(3):329-34
references.pmid: 11850619
references.citation: EMBO J. 1998 Dec 1;17(23):6979-91
references.pmid: 9843504
references.citation: Proc Natl Acad Sci U S A. 2007 Sep 4;104(36):14366-71
references.pmid: 17726103
references.citation: Mol Cell. 2014 Nov 20;56(4):580-94
references.pmid: 25457167
references.citation: Nat Protoc. 2007;2(6):1467-78
references.pmid: 17545983
references.citation: J Biol Chem. 2014 Mar 21;289(12):8353-63
references.pmid: 24515106
references.citation: Methods Mol Biol. 2002;182:85-94
references.pmid: 11768980
references.citation: Biochim Biophys Acta. 2001 May 3;1526(2):211-20
references.pmid: 11325543
references.citation: PLoS One. 2009;4(2):e4636
references.pmid: 19247482
references.citation: Genes Dev. 2004 May 1;18(9):1035-46
references.pmid: 15105378
references.citation: J Biol Chem. 2003 Jan 10;278(2):823-8
references.pmid: 12403775
references.citation: Genes Dev. 2001 Dec 15;15(24):3231-6
references.pmid: 11751628
references.citation: J Cell Biol. 1990 Mar;110(3):569-80
references.pmid: 2307700
references.citation: Mol Cell. 2011 May 6;42(3):285-96
references.pmid: 21549307
references.citation: Nature. 2007 May 24;447(7143):399-406
references.pmid: 17522672
references.citation: Science. 1993 Jun 25;260(5116):1926-8
references.pmid: 8316832
references.citation: Curr Opin Genet Dev. 2014 Apr;25:50-61
references.pmid: 24556270
references.citation: Biochem Cell Biol. 2005 Aug;83(4):548-54
references.pmid: 16094458
references.citation: Dev Dyn. 2001 May;221(1):92-105
references.pmid: 11357197
references.citation: Annu Rev Biochem. 2002;71:247-73
references.pmid: 12045097
references.citation: Nucleic Acids Res. 2006 May 31;34(10):2887-905
references.pmid: 16738128
references.citation: EMBO J. 2011 May 18;30(10):1885-95
references.pmid: 21527910
references.citation: Nature. 2000 Aug 3;406(6795):541-4
references.pmid: 10952318
references.citation: J Cell Biol. 1997 Apr 21;137(2):263-74
references.pmid: 9128241
references.citation: J Cell Sci. 2012 Nov 15;125(Pt 22):5524-34
references.pmid: 22946062
references.citation: Proc Natl Acad Sci U S A. 2010 Nov 16;107(46):19873-8
references.pmid: 20974961
references.citation: EMBO J. 2003 Oct 1;22(19):5154-62
references.pmid: 14517253
references.citation: Mol Cell. 2000 Feb;5(2):355-65
references.pmid: 10882076
references.citation: EMBO J. 1999 Apr 1;18(7):1923-38
references.pmid: 10202156
references.citation: Cancer Res. 2003 Aug 1;63(15):4677-83
references.pmid: 12907649
references.citation: Nucleus. 2011 Sep-Oct;2(5):332-8
references.pmid: 21941119
references.citation: Nucleic Acids Res. 2015 Feb 18;43(3):1444-55
references.pmid: 25578963
references.citation: Nature. 2006 Jul 6;442(7098):86-90
references.pmid: 16728976
references.citation: EMBO J. 2006 Jan 25;25(2):335-45
references.pmid: 16395332
references.citation: Bioessays. 2005 Jan;27(1):17-28
references.pmid: 15612037