Functional characterisation of Schizosaccharomyces pombe meiotic linear elements

  • Heledd Fflur Dafydd

    Student thesis: Doctor of Philosophy

    Abstract

    Sexual reproduction in eukaryotes requires the creation of haploid
    gametes from diploid cells by a specialized cell division called meiosis. Correct
    alignment and orientation of the homologues on the meta phase I plate is crucial
    for the correct reductional segregation of chromosomes during meiosis. In most
    organisms, homologue alignment is concurrent with, although distinct from, a
    process called synapsis, which is the close association between the homologues
    within the context of a proteinaceous structure formed along the entire length of
    the chromosome, called the synaptonemal complex (SC).
    The model yeast system Sehizosaeeharomyees pombe exhibits an SC-free
    meiosis. In S. pombe, proteinaceous structures termed linear elements (LinEs)
    are formed, which vary in length and are classified accordingly. A major
    component of LinEs is the Rec10 protein. Different mutations in the rec10 gene
    result in distinct defects in the formation and/or development of LinEs alongside
    varying effects on meiotic recombination.
    In this study, we demonstrate the temporal order of LinE formation and
    maturation, correlated with Rad51 and Hop1 localisation. Cytological analysis of
    Hop1 localisation revealed the presence of as yet unidentified structures.
    A multiple sequence alignment of the C-terminal domain of Rec10 with
    axial proteins of other organisms exhibited domain homology, with some
    residues being highly conserved. Based on this, this study generated several
    rec10 mutants exhibiting different point mutations within the C-terminal domain
    of rec10. In this study we analysed the temporal profile of LinE development and
    the recombination proficiency of these mutants.
    This study analysed the temporal profile of LinE development in mutants
    of LinE-associated proteins, previously demonstrated as having altered LinE
    formation; demonstrating that Rec27 is not required for initiation of LinE
    formation, as previously proposed.
    In addition, we present data that is in agreement with the regions of
    crossover preference model. We also analyse LinE formation in several rec10
    heterozygous diploids, demonstrating semi-dominance of some rec10 mutant
    alleles.
    Date of Award2011
    Original languageEnglish
    Awarding Institution
    • Bangor University

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