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Title: Structure of schizosaccharomyces pombe DNA in mouse cells
Author: McManus, John
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1993
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Integrated Schizosaccharomyces pombe (S. pombe) transgenomes in murine cells display a cytological constriction at metaphase. That the metaphase constriction is common to all integrated S. pombe transgenomes suggests that interactions between S. pombe sequences and murine proteins are the cause of these apparently aberrant structures. To investigate which aspects of chromatin packaging within the S. pombe transgenomes are related to this constriction I have studied one of the hybrid cell lines F1.1. I have analysed the amount of S. pombe DNA forming this transgenome and selected regions of it for further investigation. DNA methylation, nucleosome packaging and nucleoskeleton attachment were investigated as potential causes of the constriction. It was found that DNA methylation accumulated within the transgenome during culture, although no correlating alteration in metaphase structure was detected. Methylation was therefore felt not to be related to the observed structure of the transgenome. The transgenome was found to adopt a murine nucleosomal repeat of 185bp, rather than retain its original 160bp S. pombe repeat. Adoption of the host independent of the structure of introduced DNA. This showed that nuclesome not on the underlying DNA sequence. It was possible to demonstrate differential attachment of both murine and S. pombe sequences to the mouse nucleoskeleton. Analysis of and extended region of S. pombe DNA suggests that attachment may by closer, <20kb apart, in the S pombe transgenome, than the 80kb separation expected for mouse DNA. Packaging of these smaller loops at metaphase may lead to the observed constricted structure of the S. pombe transgenomes.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available