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. 2002 Mar;22(6):1615–1625. doi: 10.1128/MCB.22.6.1615-1625.2002

FIG. 2.

FIG. 2.

Interaction of Snf5, Swi1, and Swi2/Snf2 in the context of intact SWI/SNF with Hap4. (A) Diagram of the photo-cross-linking reagent Sulfo-SBED. (B) Schematic of the photo-cross-linking experiments. Photo-cross-linking experiments were performed with purified SWI/SNF complex and an activator that was previously conjugated with Sulfo-SBED. Where indicated, the reaction mixtures were exposed to UV light (312 nm) at 6 cm for 8 min at room temperature. The activator was separated from interacting SWI/SNF subunits upon the addition of DTT, which reduces the disulfide bond. (C) Snf5, Swi1, and Swi2/Snf2 are cross-linked by GST-Hap4. The photo-cross-linking reaction mixtures were run on an SDS-4 to 15% gradient polyacrylamide gel. The proteins were transferred to PVDF membrane and immunoblotted with streptavidin-HRP or the indicated SWI/SNF-specific antibodies. The asterisks indicate photo-cross-linked SWI/SNF subunits. (D) SWI/SNF subunits are not cross-linked by GST alone. The photo-cross-linking experiment was performed as described for panels B and C.