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Source URL: https://webapps.wesleyan.edu/grad_thesis/thesis_file/3242
Original filename: MacDonough_Gwyneth_Item_26_Copies.pdf"},{"label":"Author-supplied Keywords","value":["Epigenetics","Chromatin structure","Chromatin factor","Histone","Hho1","Htz1","Hmo1","Chromosome stability","Plasmid stability","Non-homologous end joining"]},{"label":"Degree","value":["Master of Arts"]},{"label":"Description","value":"

General chromatin factors have multiple roles in maintaining chromosome stability. We examined the influence of linker histone H1 (encoded by HHO1), variant histone H2A.Z (encoded by HTZ1), and the HMG-box protein Hmo1 on chromosome stability in S. cerevisiae. We have previously shown that loss of Htz1 in yeast cells causes defects in chromosome cohesion and condensation. A genome integrity assay indicates that cells lacking Htz1 also exhibit increases in chromosome recombination/truncation events. Cells lacking Hho1 exhibit no phenotype in this assay, but remarkably, loss of Hho1 can suppress the instability seen in cells lacking Htz1. A similar pattern is observed when assessing the stability of plasmid mini chromosomes and the efficacy of non-homologous end joining. The non-canonical HMG-box protein Hmo1 has been proposed to assume some linker histone functions in yeast cells. Interestingly, we find that yeast lacking Hmo1 also exhibit decreased stability of plasmid mini chromosomes, and this instability is also suppressed by loss of Hho1. Interestingly, we do not observe a similar antagonism between Hmo1 and Hho1 for non-homologous end joining or growth. We also examine the relationship between Hmo1 and Htz1 and demonstrate that loss of Htz1 is able to suppress defects in plasmid retention, growth, and potentially non-homologous end joining in hmo1∆ cells. The relevance of Hmo1 as a functional analog for linker histone H1 is also explored by analyzing the role of the C-terminal domain in cell proliferation and plasmid retention. We show that the CTD is important for plasmid retention, but dispensable for cell proliferation. The genetic interactions between Hho1 and Htz1 in addition to Hho1 and Hmo1 are utilized to conduct a structure-function analysis of Hho1. We demonstrate that Hho1’s second globular domain, in addition to residues K95, K96, Y47A, and P106A, make contributions to Hho1 function. Overall, these experiments indicate a balance of basic chromatin factor activities is required to preserve genome integrity.

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