Chromatin structure has been studied by many investigators to understand its role in the biological activity of DNA, namely, its role in gene expression. The focus has been on higher organisms and a great deal has been learned about the fundamental unit of chromatin, the nucleosome. This proposal involves an analysis of chromatin structure of a lower eukaryotic organism, the yeast Saccharomyces cerevisiae. There appear to be significant differences between the chromatin structure of higher and lower organisms. There are two major reasons for undertaking these studies. (1) Yeast is often used as a host for recombinant DNA studies. Optimal gene expression of foreign DNA will probably require a full understanding of yeast chromatin. (2) As a comparative study, differences in yeast and higher eukaryotic chromatin structure will be clearly defined by a number of physical and enzymatic methods. Reconstitution experiments using hybrid mixtures of histones from yeast and higher eukaryotes will provide information which will be used to assign particular changes in the chromatin structure to specific proteins (histones).
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