This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Metazoan genomes and those of many plants and fungi encode genes for histone H3 and the variant H3.3, which differ at four amino acid positions. H3.3 has garnered much interest recently because, in contrast to H3, it displays deposition at active genes outside of S phase. It has therefore been proposed that it plays a specific role in the inheritance of epigenetic states, such as those mediated by specific histone modifications. However, to our knowledge, the effect of removing H3.3 from cells has not been reported. We therefore addressed its function by taking advantage of the fact that H3 and H3.3 are each encoded by a single locus in the genetically tractable basidiomycetous fungus, Cryptococcus neoformans. As part of this effort we are using affinity purification and MudPIT to characterize the soluble deposition complexes for H3 and H3.3. Our studies have revealed perfect conservation between C. neoformans and humans of the subunits of these deposition complexes.
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