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Reference: Maiorano, Domenico., (1995). Role of cdc21+ and related genes in eukaryotic chromosome replication. DPhil. University of Oxford.Citable link to this page:

 

Role of cdc21+ and related genes in eukaryotic chromosome replication

Abstract: The Schizosaccharomyces pombe cdc21+ gene product is related to the Mcm2-3-5 family of replication proteins.By phylogeny analysis of their protein sequences and screening for cdc21+-related sequences using molecular probes I have suggested that at least six types ofcdc21+-related genes may be present in the yeast genome.The isolation of interaction suppressors of the cdc21ts mutant was attempted byoverexpression of an S. pombe cDNA library. Two cDNAs were isolated, ts11+ anddom1+, whose overexpression specifically affected the viability of cdc21ts cells undercertain conditions. The predicted dom1 protein is 60% identical to the budding yeastHMG-like Nhp2 protein.I have studied the phenotype of S. pombe cells overexpressing the cdc21+ geneand amino-terminal truncations of it. Overexpression of the cdc21+ gene caused cellelongation but cells were not significantly affected in growth rate. Cells overexpressingthe carboxyl-terminal part of cdc21+ arrested in S phase and also entered mitosis in theabsence of nuclear division.The possibility that chromosomes in cdc21ts arrested cells may be damaged wasinvestigated by pulsed field gel electrophoresis. No differences could be foundcompared to wild-type chromosomes. I have also studied the arrest phenotype of cdc21 rad1 and cdc21 cdc2.3w double mutants. Both strains entered mitosis at the restrictivetemperature indicating that cdc21ts cells arrest in S phase and may contain DNAdamage.I have generated two new mutant alleles of cdc21+. The first allele was made bydeleting most of the cdc21+ open reading frame (ORF). The second allele wasconstructed by placing the cdc21+ ORF under control a regulatable promoter. Theresulting construct was used to complement the cdc21 deletion. Both mutants wereinviable under appropriate conditions arresting in S phase as elongated cells, although aproportion of them (15-20%) entered mitosis in the absence of nuclear division.

Type of Award:DPhil Level of Award:Doctoral Awarding Institution: University of Oxford Notes:This thesis was digitised thanks to the generosity of Dr Leonard Polonsky

Bibliographic Details

Issue Date: 1995Identifiers

Urn: uuid:e4813692-f9c5-4f81-9fb8-2a13413c04bb

Source identifier: 603832171 Item Description

Type: Thesis;

Language: eng Subjects: Schizosaccharomyces pombe Genetics Cytology Tiny URL: td:603832171

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Autor: Maiorano, Domenico. - institutionUniversity of Oxford facultyDept. of Zoology - - - - Bibliographic Details Issue Date: 1995 - Id

Fuente: https://ora.ox.ac.uk/objects/uuid:e4813692-f9c5-4f81-9fb8-2a13413c04bb



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