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Title: Protein-protein interactions of the DNA polymerase ð complex in the fission yeast Schizosaccharomyces pombe
Author: Garcia, Javier Sanchez
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2003
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In eukaryotes there are three essential DNA polymerases that are involved in the bulk o f DNA replication: pola, polS and pole. P ola is involved in generating a short RNA-DNA primer. PolS and 8 are involved in the elongation process o f DNA replication. It has been suggested that polS is the key enzyme that performs all o f the processive DNA replication since the catalytic domains o f pole are not essential. In S. pombe polδ is comprised of four subunits: Pol3- the catalytic or A subunit, Cdcl- the B subunit, Cdc27- the C subunit, and Cdml- the D subunit. Polδ in S. cerevisiae and mammals have homologues o f these subunits, except for the D subunit of which there is no homologue in S. cerevisiae. In this thesis polS from S. pombe has been studied in two ways. One approach was to investigate the protein-protein interactions within polδ, and the other was to investigate Cdcl, the highly conserved B subunit of unknown function. The proteinprotein interactions were investigated using a combination of two-hybrid assays and mutational analysis. Cdcl was investigated by performing extensive mutational analysis using both random and site directed methods. The combination of approaches has demonstrated that the C- terminal ZnF2 region of both S. pom be and S. cerevisiae A subunit (Pol3) is involved in the direct binding to the B subunit. The four cysteines present in the zinc finger are involved in maintaining the structure of both S. pombe and S. cerevisiae ZnF2. Mutational analysis o f Cdcl (the S. pombe B subunit) has identified a conserved region (DomIII) that could be involved in the function of Cdcl. Additionally, binding assays with the Cdcl mutants have suggested a region of Cdcl (from amino acids 293 to 329) as being involved in the binding to Pol3.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available