Purification and characterization of mSin3A-containing Brg1 and hBrm chromatin remodeling complexes.
Alteration of nucleosomes by ATP-dependent remodeling complexes represents a critical step in the regulation of transcription. The human SWI/SNF (hSWI/SNF) family is composed of complexes that contain either Brg1 or hBrm as the central ATPase; however, these separate complexes have not been compared functionally. Here we describe the establishment of cell lines that express epitope-tagged Brg1 and hBrm and a characterization of the complexes associated with these two ATPases. We show that Brg1 fractionates into two complexes that differ in activity and subunit composition, whereas hBrm is found in one complex with lower activity than the Brg1 complexes. These three complexes can remodel nucleosomal arrays, increase restriction enzyme accessibility, and hydrolyze ATP in a DNA-dependent manner. The three complexes differ markedly in their ability to remodel mononucleosomal core particles. We also show that the hBrm complex and one of the Brg1 complexes contain components of the mammalian Sin3 (mSin3) complex. In addition, we have found that Brg1, hBrm, and BAF155 can interact specifically with mSin3A in vitro, showing a direct association of hSWI/SNF complexes with proteins involved in gene repression. These unexpected functional characteristics indicate that these hSWI/SNF complexes play diverse regulatory roles.
Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA. Sif.1@osu.edu
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Sif
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Saurin
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Imbalzano
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Kingston
D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism*; D004265:DNA Helicases; D000939:Epitopes; D006655:Histone Deacetylases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; D009687:Nuclear Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D056264:Sin3 Histone Deacetylase and Corepressor Complex; D014157:Transcription Factors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D002843:Chromatin; D000939:Epitopes; D009687:Nuclear Proteins; D009707:Nucleosomes; C084011:SMARCA2 protein, human; D014157:Transcription Factors; D000255:Adenosine Triphosphate; D006655:Histone Deacetylases; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; D056264:Sin3 Histone Deacetylase and Corepressor Complex; D000251:Adenosine Triphosphatases; C084108:SMARCA4 protein, human; D004265:DNA Helicases