Investigation of the Effect of Altering Residues on H2A.Z Deposition in Saccharomyces cerevisiae

There are numerous ways in which chromatin structure can be changed. Two of these mechanisms involve alteration of histones through post-translational modifications and additions of histone variants. These variants differ from canonical histones through their structure and function. The most abundant variant, H2A.Z, is a specialized form of the canonical histone H2A. H2A.Z containing nucleosomes are located near the telomeres of chromosomes. We believe that on this histone, the threonine 87 residue is phosphorylated post-translationally and has biological functionality. As a consequence of telomeric silencing, the ADE2 gene may or may not be expressed. We plan to use placement of ADE2 gene to test if telomeric silencing is abrogated in our mutant. This is associated to the placement and functionality of the H2A.Z histone variant. Both ChIP and telomeric silencing assays are used to detect the presence and placement of H2A.Z containing nucleosomes in T87 mutants of Saccharomyces cerevisiae