An Ikaros-containing chromatin-remodeling complex in adult-type erythroid cells

D. W. O'Neill, S. S. Schoetz, R. A. Lopez, M. Castle, L. Rabinowitz, E. Shor, D. Krawchuk, M. G. Goll, M. Renz, H. P. Seelig, S. Han, R. H. Seong, S. D. Park, T. Agalioti, N. Munshi, D. Thanos, H. Erdjument-Bromage, P. Tempst, A. Bank

Research output: Contribution to journalArticle

132 Citations (Scopus)

Abstract

We have previously described a SWI/SNF-related protein complex (PYR complex) that is restricted to definitive (adult-type) hematopoietic cells and that specifically binds DNA sequences containing long stretches of pyrimidines. Deletion of an intergenic DNA-binding site for this complex from a human β-globin locus construct results in delayed human γ- to β-globin switching in transgenic mice, suggesting that the PYR complex acts to facilitate the switch. We now show that PYR complex BNA-binding activity also copurifies with subunits of a second type of chromatin-remodeling complex, nucleosome-remodeling deacetylase (NuRD), that has been shown to have both nucleosome-remodeling and histone deacetylase activities. Gel supershift assays using antibodies to the ATPase-helicase subunit of the NuRD complex, Mi-2 (CHD4), confirm that Mi-2 is a component of the PYR complex. In addition, we show that the hematopoietic cell-restricted zinc finger protein Ikaros copurifies with PYR complex DNA-binding activity and that antibodies to Ikaros also supershift the complex. We also show that NuRD and SWI/SNF components coimmunopurify with each other as well as with Ikaros. Competition gel shift experiments using partially purified PYR complex and recombinant Ikaros protein indicate that Ikaros functions as a DNA-binding subunit of the PYR complex. Our results suggest that Ikaros targets two types of chromatin-remodeling factors - activators (SWI/SNF) and repressors (NuRD) - in a single complex (PYR complex) to the β-globin locus in adult erythroid cells. At the time of the switch from fetal to adult globin production, the PYR complex is assembled and may function to repress γ-globin gene expression and facilitate β- to β-globin switching.

Original languageEnglish (US)
Pages (from-to)7572-7582
Number of pages11
JournalMolecular and Cellular Biology
Volume20
Issue number20
DOIs
StatePublished - 2000

Fingerprint

Erythroid Cells
Chromatin Assembly and Disassembly
Globins
Nucleosomes
Mi-2 Nucleosome Remodeling and Deacetylase Complex
Ikaros Transcription Factor
Gels
Pyrimidines
Intergenic DNA
Histone Deacetylases
Antibodies
DNA
Zinc Fingers
Recombinant Proteins
Transgenic Mice
Adenosine Triphosphatases
Binding Sites
Gene Expression
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

O'Neill, D. W., Schoetz, S. S., Lopez, R. A., Castle, M., Rabinowitz, L., Shor, E., ... Bank, A. (2000). An Ikaros-containing chromatin-remodeling complex in adult-type erythroid cells. Molecular and Cellular Biology, 20(20), 7572-7582. https://doi.org/10.1128/MCB.20.20.7572-7582.2000

An Ikaros-containing chromatin-remodeling complex in adult-type erythroid cells. / O'Neill, D. W.; Schoetz, S. S.; Lopez, R. A.; Castle, M.; Rabinowitz, L.; Shor, E.; Krawchuk, D.; Goll, M. G.; Renz, M.; Seelig, H. P.; Han, S.; Seong, R. H.; Park, S. D.; Agalioti, T.; Munshi, N.; Thanos, D.; Erdjument-Bromage, H.; Tempst, P.; Bank, A.

In: Molecular and Cellular Biology, Vol. 20, No. 20, 2000, p. 7572-7582.

Research output: Contribution to journalArticle

O'Neill, DW, Schoetz, SS, Lopez, RA, Castle, M, Rabinowitz, L, Shor, E, Krawchuk, D, Goll, MG, Renz, M, Seelig, HP, Han, S, Seong, RH, Park, SD, Agalioti, T, Munshi, N, Thanos, D, Erdjument-Bromage, H, Tempst, P & Bank, A 2000, 'An Ikaros-containing chromatin-remodeling complex in adult-type erythroid cells', Molecular and Cellular Biology, vol. 20, no. 20, pp. 7572-7582. https://doi.org/10.1128/MCB.20.20.7572-7582.2000
O'Neill, D. W. ; Schoetz, S. S. ; Lopez, R. A. ; Castle, M. ; Rabinowitz, L. ; Shor, E. ; Krawchuk, D. ; Goll, M. G. ; Renz, M. ; Seelig, H. P. ; Han, S. ; Seong, R. H. ; Park, S. D. ; Agalioti, T. ; Munshi, N. ; Thanos, D. ; Erdjument-Bromage, H. ; Tempst, P. ; Bank, A. / An Ikaros-containing chromatin-remodeling complex in adult-type erythroid cells. In: Molecular and Cellular Biology. 2000 ; Vol. 20, No. 20. pp. 7572-7582.
@article{20861e83b2cd4f3a9d1afcb93f4288f6,
title = "An Ikaros-containing chromatin-remodeling complex in adult-type erythroid cells",
abstract = "We have previously described a SWI/SNF-related protein complex (PYR complex) that is restricted to definitive (adult-type) hematopoietic cells and that specifically binds DNA sequences containing long stretches of pyrimidines. Deletion of an intergenic DNA-binding site for this complex from a human β-globin locus construct results in delayed human γ- to β-globin switching in transgenic mice, suggesting that the PYR complex acts to facilitate the switch. We now show that PYR complex BNA-binding activity also copurifies with subunits of a second type of chromatin-remodeling complex, nucleosome-remodeling deacetylase (NuRD), that has been shown to have both nucleosome-remodeling and histone deacetylase activities. Gel supershift assays using antibodies to the ATPase-helicase subunit of the NuRD complex, Mi-2 (CHD4), confirm that Mi-2 is a component of the PYR complex. In addition, we show that the hematopoietic cell-restricted zinc finger protein Ikaros copurifies with PYR complex DNA-binding activity and that antibodies to Ikaros also supershift the complex. We also show that NuRD and SWI/SNF components coimmunopurify with each other as well as with Ikaros. Competition gel shift experiments using partially purified PYR complex and recombinant Ikaros protein indicate that Ikaros functions as a DNA-binding subunit of the PYR complex. Our results suggest that Ikaros targets two types of chromatin-remodeling factors - activators (SWI/SNF) and repressors (NuRD) - in a single complex (PYR complex) to the β-globin locus in adult erythroid cells. At the time of the switch from fetal to adult globin production, the PYR complex is assembled and may function to repress γ-globin gene expression and facilitate β- to β-globin switching.",
author = "O'Neill, {D. W.} and Schoetz, {S. S.} and Lopez, {R. A.} and M. Castle and L. Rabinowitz and E. Shor and D. Krawchuk and Goll, {M. G.} and M. Renz and Seelig, {H. P.} and S. Han and Seong, {R. H.} and Park, {S. D.} and T. Agalioti and N. Munshi and D. Thanos and H. Erdjument-Bromage and P. Tempst and A. Bank",
year = "2000",
doi = "10.1128/MCB.20.20.7572-7582.2000",
language = "English (US)",
volume = "20",
pages = "7572--7582",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "20",

}

TY - JOUR

T1 - An Ikaros-containing chromatin-remodeling complex in adult-type erythroid cells

AU - O'Neill, D. W.

AU - Schoetz, S. S.

AU - Lopez, R. A.

AU - Castle, M.

AU - Rabinowitz, L.

AU - Shor, E.

AU - Krawchuk, D.

AU - Goll, M. G.

AU - Renz, M.

AU - Seelig, H. P.

AU - Han, S.

AU - Seong, R. H.

AU - Park, S. D.

AU - Agalioti, T.

AU - Munshi, N.

AU - Thanos, D.

AU - Erdjument-Bromage, H.

AU - Tempst, P.

AU - Bank, A.

PY - 2000

Y1 - 2000

N2 - We have previously described a SWI/SNF-related protein complex (PYR complex) that is restricted to definitive (adult-type) hematopoietic cells and that specifically binds DNA sequences containing long stretches of pyrimidines. Deletion of an intergenic DNA-binding site for this complex from a human β-globin locus construct results in delayed human γ- to β-globin switching in transgenic mice, suggesting that the PYR complex acts to facilitate the switch. We now show that PYR complex BNA-binding activity also copurifies with subunits of a second type of chromatin-remodeling complex, nucleosome-remodeling deacetylase (NuRD), that has been shown to have both nucleosome-remodeling and histone deacetylase activities. Gel supershift assays using antibodies to the ATPase-helicase subunit of the NuRD complex, Mi-2 (CHD4), confirm that Mi-2 is a component of the PYR complex. In addition, we show that the hematopoietic cell-restricted zinc finger protein Ikaros copurifies with PYR complex DNA-binding activity and that antibodies to Ikaros also supershift the complex. We also show that NuRD and SWI/SNF components coimmunopurify with each other as well as with Ikaros. Competition gel shift experiments using partially purified PYR complex and recombinant Ikaros protein indicate that Ikaros functions as a DNA-binding subunit of the PYR complex. Our results suggest that Ikaros targets two types of chromatin-remodeling factors - activators (SWI/SNF) and repressors (NuRD) - in a single complex (PYR complex) to the β-globin locus in adult erythroid cells. At the time of the switch from fetal to adult globin production, the PYR complex is assembled and may function to repress γ-globin gene expression and facilitate β- to β-globin switching.

AB - We have previously described a SWI/SNF-related protein complex (PYR complex) that is restricted to definitive (adult-type) hematopoietic cells and that specifically binds DNA sequences containing long stretches of pyrimidines. Deletion of an intergenic DNA-binding site for this complex from a human β-globin locus construct results in delayed human γ- to β-globin switching in transgenic mice, suggesting that the PYR complex acts to facilitate the switch. We now show that PYR complex BNA-binding activity also copurifies with subunits of a second type of chromatin-remodeling complex, nucleosome-remodeling deacetylase (NuRD), that has been shown to have both nucleosome-remodeling and histone deacetylase activities. Gel supershift assays using antibodies to the ATPase-helicase subunit of the NuRD complex, Mi-2 (CHD4), confirm that Mi-2 is a component of the PYR complex. In addition, we show that the hematopoietic cell-restricted zinc finger protein Ikaros copurifies with PYR complex DNA-binding activity and that antibodies to Ikaros also supershift the complex. We also show that NuRD and SWI/SNF components coimmunopurify with each other as well as with Ikaros. Competition gel shift experiments using partially purified PYR complex and recombinant Ikaros protein indicate that Ikaros functions as a DNA-binding subunit of the PYR complex. Our results suggest that Ikaros targets two types of chromatin-remodeling factors - activators (SWI/SNF) and repressors (NuRD) - in a single complex (PYR complex) to the β-globin locus in adult erythroid cells. At the time of the switch from fetal to adult globin production, the PYR complex is assembled and may function to repress γ-globin gene expression and facilitate β- to β-globin switching.

UR - http://www.scopus.com/inward/record.url?scp=0033805828&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033805828&partnerID=8YFLogxK

U2 - 10.1128/MCB.20.20.7572-7582.2000

DO - 10.1128/MCB.20.20.7572-7582.2000

M3 - Article

VL - 20

SP - 7572

EP - 7582

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 20

ER -