Deducing the post-transcriptional role of the Pumilio protein Puf3 in fission yeast.

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dc.contributor.advisorChua, Gordon
dc.contributor.authorGeorge, Iain Fraser Scotney
dc.contributor.committeememberMuench, Douglas G.
dc.contributor.committeememberTurner, Raymond Joseph
dc.contributor.committeememberGrewal, Savraj S.
dc.contributor.committeememberKothe, Ute
dc.date2019-11
dc.date.accessioned2019-06-03T18:32:54Z
dc.date.available2019-06-03T18:32:54Z
dc.date.issued2019-05-29
dc.description.abstractPumilio family proteins (Pufs) are a highly-conserved group of post-transcriptional regulators in eukaryotes. Puf proteins contain an arc-shaped Pumilio homology domain with several coiled repeats that bind to potentially predictable sequences in the 3’-UTR of their mRNA targets. The binding of Puf proteins to mRNA targets can influence their stability, translation and cellular localization. In S. pombe, there are seven non-essential genes that encode classical Puf proteins. The biological function of these Puf proteins and their mRNA targets remain unknown. In this study, we investigated the biological role and mRNA targets of Puf3 in S. pombe. Over-expression of puf3+ with the nmt41 promoter results in reduced fitness and an enrichment of elongated cells arrested in mitosis. Phenotypic characterization of puf3 overexpression strains and genetic interaction screens identified a cell cycle regulatory role for Puf3, principally at the G2/M transition and possibly at the G1/S control point. Genetic interaction screening, fluorescence microscopy and tandem mass spectrometry analysis of immunoprecipitated Puf3-HA identified Puf3-interacting proteins in the post-transcriptional machinery of 3’-deadenylation of mRNA and localized to stress granules. Bioinformatic analysis of the Puf proteins in S. pombe identified the Puf repeat domains and these were used to enumerate the predicted Puf binding element in the transcriptome. The enumeration data and annotation data were used to identify putative mRNA targets from microarray, RNA-seq and RNA-immunoprecipitation sequencing experiments. These identified several components of the kinetochore and mitotic checkpoint as putative mRNA targets of Puf3.en_US
dc.identifier.citationGeorge, I. F. S. (2019). Deducing the post-transcriptional role of the Pumilio protein Puf3 in fission yeast (Doctoral thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.en_US
dc.identifier.doihttp://dx.doi.org/10.11575/PRISM/36616
dc.identifier.urihttp://hdl.handle.net/1880/110465
dc.language.isoengen_US
dc.publisher.facultyScienceen_US
dc.publisher.institutionUniversity of Calgaryen
dc.rightsUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.en_US
dc.subjectPumilioen_US
dc.subjectPuf3en_US
dc.subjectmRNAen_US
dc.subjectDeadenylationen_US
dc.subjectSchizosaccharomyces pombeen_US
dc.subjectFission Yeasten_US
dc.subject.classificationBiologyen_US
dc.subject.classificationBioinformaticsen_US
dc.subject.classificationBiology--Cellen_US
dc.subject.classificationGeneticsen_US
dc.subject.classificationMicrobiologyen_US
dc.subject.classificationBiology--Molecularen_US
dc.titleDeducing the post-transcriptional role of the Pumilio protein Puf3 in fission yeast.en_US
dc.typedoctoral thesisen_US
thesis.degree.disciplineBiological Sciencesen_US
thesis.degree.grantorUniversity of Calgaryen_US
thesis.degree.nameDoctor of Philosophy (PhD)en_US
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