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Title: Probing Translational Control during Drosophila Oogenesis: from nanos to the Entire Maternal Transcriptome
Authors: Peng, Yingshi
Advisors: Gavis, Elizabeth R
Contributors: Molecular Biology Department
Keywords: Fragile X mental retardation protein (FMRP)
Post-transcriptional gene regulation
Translation elongation regulation
Translational control
Subjects: Molecular biology
Issue Date: 2021
Publisher: Princeton, NJ : Princeton University
Abstract: Translational control provides a key mechanism for the spatial and temporal regulation of eukaryotic gene expression. It plays a particularly important role in early embryonic development in organisms that rely heavily on maternally supplied mRNAs. The Drosophila developing oocyte serves as a valuable model for studying translational control. The Drosophila posterior determinant, nanos (nos), is translationally repressed throughout the oocyte cytoplasm except at the posterior pole. A multi-functional RNA-binding protein Glorund (Glo), the homolog of the mammalian hnRNP F/H family of proteins, represses translation of nos during oogenesis by targeting both translation initiation and translation elongation. To elucidate the molecular mechanism by which Glo regulates nos, I identified dFMRP as a Glo-interacting protein. By biochemically dissecting repression of nos translation in vitro, I demonstrated that dFMRP specifically inhibits translation elongation. Furthermore, I combined mutational analysis and in vivo and in vitro binding assays to show that Glo’s qRRM2 domain specifically and directly interacts with dFMRP, suggesting that Glo’s RNA-binding domains can also function as protein-protein interaction interfaces critical for its regulatory functions. Additionally, I applied ribosome footprint profiling to the Drosophila ovary at different developmental stages to identify maternal transcripts regulated during the translation elongation phase. A footprint peak-finding tool has been developed to detect transcriptome-wide ribosome stalling sites. My preliminary results suggest that developmentally regulated ribosome stalling sites may be widely present on Drosophila maternal transcripts. An A-/P-site mapping algorithm is currently under development to facilitate mechanistic analysis of ribosome stalling during oogenesis.
Alternate format: The Mudd Manuscript Library retains one bound copy of each dissertation. Search for these copies in the library's main catalog:
Type of Material: Academic dissertations (Ph.D.)
Language: en
Appears in Collections:Molecular Biology

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