title: Influence of combinatorial histone modifications on antibody and effector protein recognition.
abstract: Increasing evidence suggests that histone posttranslational modifications (PTMs) function in a combinatorial fashion to regulate the diverse activities associated with chromatin. Yet how these patterns of histone PTMs influence the adapter proteins known to bind them is poorly understood. In addition, how histone-specific antibodies are influenced by these same patterns of PTMs is largely unknown. Here we examine the binding properties of histone-specific antibodies and histone-interacting proteins using peptide arrays containing a library of combinatorially modified histone peptides. We find that modification-specific antibodies are more promiscuous in their PTM recognition than expected and are highly influenced by neighboring PTMs. Furthermore, we find that the binding of histone-interaction domains from BPTF, CHD1, and RAG2 to H3 lysine 4 trimethylation is also influenced by combinatorial PTMs. These results provide further support for the histone code hypothesis and raise specific concerns with the quality of the currently available modification-specific histone antibodies.
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC 27599, USA.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Fuchs
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Krajewski
authors.forename: Richard W
authors.initials: RW
authors.lastname: Baker
authors.forename: Victoria L
authors.initials: VL
authors.lastname: Miller
authors.forename: Brian D
authors.initials: BD
authors.lastname: Strahl
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies / Q000378:metabolism*; D000915:Antibody Affinity; D006657:Histones / Q000276:immunology* / Q000378:metabolism*; D008745:Methylation; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D017434:Protein Structure, Tertiary
chemical_list: D000906:Antibodies; D006657:Histones