Mapping Protein–DNA and Protein–Protein Interactions of ATP-Dependent Chromatin Remodelers
Chromatin plays a key regulatory role in several DNA-dependent processes as it regulates DNA access to different protein factors. Several multisubunit protein complexes interact, modify, or mobilize nucleo-somes: the basic unit of chromatin, from its original location in an ATP-dependent manner to facilitate processes, such as transcription, replication, repair, and recombination. Knowledge of the interactions of chromatin remodelers with nucleosomes is a crucial requirement to understand the mechanism of chromatin remodeling. Here, we describe several methods to analyze the interactions of multisubunit chromatin-remodeling enzymes with nucleosomes.
Key wordsNucleosome Chromatin remodeling SWI/SNF ISW2 DNA–protein interactions DNA and histone cross-linking DNA footprinting
- 13.Shen, F., Triezenberg, S. J., Hensley, P., Porter, D. and Knutson, J. R. (1996) Critical amino acids in the transcriptional activation domain of the herpesvirus protein VP16 are solvent-exposed in highly mobile protein segments. An intrinsic fluorescence study. J Biol Chem. 271 4819–4826.Google Scholar
- 18.Lannutti, B. J., Persinger, J. and Bartholomew, B. (1996) Probing the protein-DNA contacts of a yeast RNA polymerase III transcription complex in a crude extract: solid phase synthesis of DNA photoaffinity probes containing a novel photoreactive deoxycytidine analog. Biochemistry. 35 9821–9831.PubMedCrossRefGoogle Scholar
- 24.Chizzonite, R., Truitt, T., Desai, B. B., Nunes, P., Podlaski, F. J., Stern, A. S. and Gately, M. K. (1992) IL-12 receptor. I. Characterization of the receptor on phytohemagglutinin-activated human lymphoblasts. J Immunol. 148 3117–3124.Google Scholar
- 25.Chizzonite, R., Truitt, T., Podlaski, F. J., Wolitzky, A. G., Quinn, P. M., Nunes, P., Stern, A. S. and Gately, M. K. (1991) IL-12: monoclonal antibodies specific for the 40-kDa subunit block receptor binding and biologic activity on activated human lymphoblasts. J Immunol. 147 1548–1556.PubMedGoogle Scholar