{"@context":"http://iiif.io/api/presentation/2/context.json","@type":"sc:Manifest","@id":"https://iiif.quartexcollections.com/wesleyan/iiif/cbd62f0f-4e7c-4312-b2a2-f2976cff8654/manifest","label":"m2021-m681","metadata":[{"label":"Title","value":"A grave affair: investigating the unique spore lethality phenotype of the fpr3∆ zip3∆ double mutant"},{"label":"Subject","value":["Genetic recombination","Chromosome replication"]},{"label":"Rights","value":["In Copyright"]},{"label":"Repository","value":["University Archives"]},{"label":"Language","value":["English"]},{"label":"Item Information","value":"<a href=\"https://onesearch.wesleyan.edu/permalink/01CTW_WU/12cbh08/alma9932796976603768\" target=\"_blank\">M2021 M681</a>"},{"label":"Held By","value":"Wesleyan University Library, Wesleyan University, Middletown, CT, USA"},{"label":"Group","value":["Theses & Dissertations"]},{"label":"Name(s)","value":["Mitchell, Mackenzie Augusta (creator)"]},{"label":"Administrative Note","value":"Node ID: 17047<br>Author Contact: mmitchell01@wesleyan.edu<br>Local Identifier: Mitchell_Mackenzie_Item_6_2Copies<br>PID: ir:3023"},{"label":"Degree","value":["Master of Arts"]},{"label":"Description","value":"<p>Homolog pairing, synaptonemal complex (SC) assembly, and crossovers are processes essential for meiotic chromosome segregation. Errors in these pathways often lead to nondisjunction of chromosomes, resulting in aneuploid and often inviable gametes. We aim to achieve a wider understanding of the meiotic players important for successful segregation. The prolyl isomerase, Fpr3, and the E3 SUMO ligase, Zip3, were previously identified in budding yeast as proteins that act together in a checkpoint-like manner to ensure that SC assembly is dependent upon recombination initiation and homolog pairing. Interestingly, the spore viability of the fpr3Δ zip3Δ mutant is approximately twice as low as zip3Δ single mutants and other typical meiotic mutants with reduced crossovers. Here we investigate the molecular underpinnings of the fpr3Δ zip3Δ genetic interaction. We found that the spo13Δ mutation suppresses the spore lethality phenotype of fpr3Δ zip3Δ, suggesting that this synergistic phenotype relies on progression through the meiosis I division. However, we found no evidence for massive chromosome mis-segregation in the nuclei of spore products from fpr3Δ zip3Δ meiotic cells. The zip1[Δ21-163] allele also restores viability to the fpr3Δ zip3Δ mutant, indicating that the Zip1 protein exerts a toxic effect on meiosis when Fpr3 and Zip3 proteins are missing. Zip1 protein does not noticeably persist either in SC or in pairing centromeres when Fpr3 and Zip3 are missing. Our analysis of genetic mutants furthermore indicates that the extreme spore death phenotype of the fpr3Δ zip3Δ mutant is not linked to Zip1's role in promoting Msh4 crossovers, SC assembly, or centromere pairing per se. Structure-function analysis reveals that Fpr3's prolyl isomerase activity may be dispensable for its function in synergistically reigning in Zip1's toxic behavior, but Zip3's SUMOylation activity appears to be critical for its role in this process. Interestingly, removing either Mcm21 or Ctf19, two non-essential centromere-associated kinetochore proteins, imitates the catastrophic fpr3Δ zip3Δ spore lethality phenotype, but our genetic analysis indicates that these proteins act via an independent pathway. We propose a model whereby Fpr3 and Zip3 together regulate Zip1 behavior at crossover recombination sites, in a manner that affects the stability of cohesin on the arms of chromosomes. We suggest that this pathway acts in parallel to processes at the kinetochore that stabilize cohesin near centromeres, and together these processes ensure that the residual recombinant chromatids (crossover chromosomes) in fpr3Δ zip3Δ meiocytes are engaged in a stable linkage between homologous chromosomes, thus ensuring the segregation of these chromosomes on the meiosis I spindle. Smaller chromosomes (which sustain fewer crossovers than larger chromosomes) may be disproportionately affected by the absence of the Fpr3-Zip3 pathway.</p>"},{"label":"Digital Identifier","value":"m2021-m681"},{"label":"Thesis Advisor(s)","value":["MacQueen, Amy"]},{"label":"Digital Object Identifier (DOI)","value":"https://doi.org/10.14418/wes01.2.310"},{"label":"Discipline","value":["Molecular Biology and Biochemistry"]},{"label":"Date","value":"January 22, 2021"},{"label":"EDTF Date","value":"D:00 M:00 Y:2021"},{"label":"Extent","value":"viii, 148 pages : illustrations ; 28 cm"},{"label":"File Format","value":["application/pdf"]},{"label":"Genre","value":["masters theses"]},{"label":"Digital Origin","value":["born digital"]},{"label":"Type","value":["Text"]}],"description":"A grave affair: investigating the unique spore lethality phenotype of the fpr3∆ zip3∆ double 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