Early embryonic development in vertebrates is directed in part by maternal mRNAs
expressed in oocytes and stored in cytoplasmic messenger ribonucleoprotein particles (mRNPs).
Abundant evidence demonstrates the importance of mRNPs in embryonic development and in
post-embryonic cellular function; however their characterization has been hampered by lack of
suitable methodologies. The Xenopus oocyte has been the primary model system for studies of
mRNPs. YY1 is a well-studied transcriptional regulatory factor that is sequestered in the oocyte cytoplasm and present entirely in cytoplasmic oocyte mRNPs. The objective of this thesis was to examine the biochemistry of YY1 association with maternal mRNA molecules in order to shed light on the role of YY1 in development and the poorly understood biology of oocyte mRNPs.
The initial working hypotheses were that association of YY1 with mRNPs is dependent on
sequence-specific RNA-binding activity and, therefore, that YY1 associates with a definite
subset of maternal mRNA. A number of unique methods were developed in this study to address
these hypotheses. RNA immunoprecipitation-DNA microarray (RIP-CHIP) analysis establishes
that YY1 associates with a subset of mRNAs in the oocyte pool. A novel sequence-specific
RNA-binding activity of the YY1 protein is demonstrated, and the RNA-binding activity of YY1
is shown to be required for its association with oocyte mRNPs in vivo. The functional roles of
YY1 mRNA substrates are discussed in the context of embryological development and the
biological function of YY1 in oocyte mRNPs. Extension of the experimental approaches
developed in this thesis to the entire set of mRNP proteins would significantly advance our
understanding of mRNP composition and heterogeneity, as well as the biological function of maternal mRNAs and mRNPs in development.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:SSU.etd-11012010-125628 |
| Date | 01 March 2011 |
| Creators | Belak, Zachery Roderick |
| Contributors | Damjanovski, Sashko, Harkness, Troy, Schreyer, David, Kulyk, William, Chelico, Linda, Ovsenek, Nick |
| Publisher | University of Saskatchewan |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://library.usask.ca/theses/available/etd-11012010-125628/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Saskatchewan or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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