Swi3p controls SWI/SNF assembly and ATP-dependent H2A-H2B displacement

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Abstract

Yeast SWI/SNF is a multisubunit, 1.14-MDa ATP-dependent chromatin-remodeling enzyme required for transcription of a subset of inducible genes. Biochemical studies have demonstrated that SWI/SNF uses the energy from ATP hydrolysis to generate superhelical torsion, mobilize mononucleosomes, enhance the accessibility of nucleosomal DNA and remove H2A-H2B dimers from mononucleosomes. Here we describe the ATP-dependent activities of a SWI/SNF sub complex that is composed of only three subunits, Swi2p, Arp7p and Arp9p. Whereas this sub complex is fully functional in most remodeling assays, Swi2p–Arp7p–Arp9p is defective for ATP-dependent removal of H2A-H2B dimers. We identify the acidic N terminus of the Swi3p subunit as a novel H2A-H2B–binding domain required for ATP-dependent dimer loss. Our data indicate that H2A-H2B dimer loss is not an obligatory consequence of ATP-dependent DNA translocation, and furthermore they suggest that SWI/SNF is composed of at least four interdependent modules.

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Figure 1: The SANT domain of Swi3p is required for SWI/SNF assembly.
Figure 2: Swi2–Arp7–Arp9 sub complex has ATPase activity and can generate superhelical torsion.
Figure 3: Chromatin-remodeling activity of the minimal Swi2p–Arp7p–Arp9p sub complex.
Figure 4: Swi2–Arp7–Arp9 sub complex is defective in catalysis of histone H2A-H2B dimer displacement.
Figure 5: Swi3p has a novel histone-binding domain.
Figure 6: The acidic N-terminal domain of Swi3p is required for displacement of histone H2A.
Figure 7: A proposed model for SWI/SNF remodeling and functional organization.

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Acknowledgements

We thank J. Reese (Penn State University) for SWI/SNF antibodies used for western blotting, and T. Owen-Hughes (Dundee) for the gift of T4 endonuclease VII. This work was supported by a grant from the US National Institutes of Health to C.L.P.

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Correspondence to Craig L Peterson.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Mononucleosome mobility assay. (PDF 52 kb)

Supplementary Fig. 2

601 mononucleosome remodeling assays. (PDF 65 kb)

Supplementary Methods (PDF 59 kb)

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Yang, X., Zaurin, R., Beato, M. et al. Swi3p controls SWI/SNF assembly and ATP-dependent H2A-H2B displacement. Nat Struct Mol Biol 14, 540–547 (2007) doi:10.1038/nsmb1238

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