title: Analysis of nuclear receptor acetylation.
abstract: Acetylation is an essential post-translational modification in which an acetyl group is covalently conjugated to a protein substrate. Histone acetylation was first proposed nearly half a century ago by Dr. Vincent Allfrey. Subsequent studies have shown that acetylated core histones are often associated with transcriptionally active chromatin. Acetylation at lysine residues of histone tails neutralizes the positive charge, which decreases the binding ability to DNA and increases the accessibility of transcription factors and co-activators to the chromatin template. In addition to histones, a number of non-histone substrates are acetylated. Acetylation of non-histone proteins governs biological processes, including cellular proliferation and survival, transcriptional activity, and intracellular trafficking. We demonstrated that acetylation of transcription factors can regulate cellular growth. Further, we have shown that nuclear receptors are acetylated at a phylogenetically conserved motif. Since our initial observations with the estrogen and androgen receptors, more than a dozen nuclear receptors have been shown to function as substrates for acetyltransferases with a variety of new methods (Fig. 11.1). This chapter focuses on the protocol used in the studies of NR acetylation and de-acetylation. We will discuss the potential pitfalls of each method.
authors.affiliation: Department of Stem Cell Biology and Regenerative Medicine, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA. chenguang.wang@jefferson.edu
authors.forename: Chenguang
authors.initials: C
authors.lastname: Wang
authors.forename: Michael
authors.initials: M
authors.lastname: Powell
authors.forename: Lifeng
authors.initials: L
authors.lastname: Tian
authors.forename: Richard G
authors.initials: RG
authors.lastname: Pestell
mesh_terms: D000107:Acetylation; D001681:Biological Assay; D018929:Cell Culture Techniques; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D010802:Phylogeny; D040901:Proteomics; D018160:Receptors, Cytoplasmic and Nuclear / Q000145:classification / Q000235:genetics / Q000378:metabolism*
chemical_list: D006657:Histones; D018160:Receptors, Cytoplasmic and Nuclear