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Structural analysis of the yeast SWI/SNF chromatin remodeling complex

Abstract

Elucidating the mechanism of ATP-dependent chromatin remodeling is one of the largest challenges in the field of gene regulation. One of the missing pieces in understanding this process is detailed structural information on the enzymes that catalyze the remodeling reactions. Here we use a combination of subunit radio-iodination and scanning transmission electron microscopy to determine the subunit stoichiometry and native molecular weight of the yeast SWI/SNF complex. We also report a three-dimensional reconstruction of yeast SWI/SNF derived from electron micrographs.

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Figure 1: Swi2p was immunoprecipitated from whole cell extracts made from yeast strains CY831 (SWI2-HA3), CY832 (SWI2-Myc18) and CY889 (SWI2-Myc18/SWI2-HA3).
Figure 2: Purification and stoichiometry of the SWI/SNF complex.
Figure 3: STEM mass analysis of individual SWI/SNF complexes reveals a unimodal distribution with a mean of 1.14 MDa.
Figure 4: 3D structure of the yeast SWI/SNF complex.
Figure 5: Principal features of the SWI/SNF reconstruction shown in Fig. 4.

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Acknowledgements

We thank B. Cairns for communicating the unpublished sequence of SWP82, D. Kelleher for help with the radio-iodination studies, S.J. Ludtke for assistance with EMAN and J.S. Wall and M. Simon of the Brookhaven National Laboratory STEM Facility for mass determination. The STEM is an NIH Research Resource also supported by DOE and OBER. These studies were supported by NIH grants to C.L.W. and to C.L.P.

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Correspondence to Craig L. Peterson.

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Smith, C., Horowitz-Scherer, R., Flanagan, J. et al. Structural analysis of the yeast SWI/SNF chromatin remodeling complex. Nat Struct Mol Biol 10, 141–145 (2003). https://doi.org/10.1038/nsb888

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