UBC Theses and Dissertations
A quest for function : identification of proteins that interact with the Drosophila melanogaster histone variant H2A.v.D McNamee, Jason Allan
Chromatin is a structure within the cell composed of DNA , histone and non-histone proteins. It packages the cells genome and thus regulates gene expression throughout development. Histones are an important component of chromatin, forming the nucleosome structure on which the DNA wraps in order to be condensed and packaged. Histone variants are variations of these histones that are encoded by single copy genes located elsewhere in the genome. Most of these variants are essential to the organism and cannot be compensated for by their core counter parts and therefore must have a defined role in the cell. One of the most studied classes of these variants is the H2A.Z class, a variant class of the major H2A core histone. Homo logs have been found in a variety of organisms from yeast to mammals all exhibiting very high sequence conservation. Here we attempted to determine function of one of these homologs of H2A.Z, Drosophila H2A.vD. We attempted to gain an understanding of protein function by identifying proteins with which H2A.vD interacts. We employed a Protein Interaction Trap direct test method followed by GST pulldowns in order to confirm physical direct interactions. We identified the Drosophila gene CG5515 as a direct interactor o f H2A.vD in vitro. We used genetic analysis in an attempt to understand the in vivo significance of the interaction. We believe that the hypothetical protein product of CG5515 may work in a phosphorylation pathway for H2A.vD. This modification may either aid H2A.vD in generating alternate chromatin structure or possibly in its response to DNA damage.
Item Citations and Data