BOKU - Universität für Bodenkultur Wien - Forschungsinformationssystem

Gewählte Diplomarbeit / Masterarbeit:

Shady Younis (2011): Molecular Characterization of ZC3H11A Functions and Expression Patterns During Embryonic Development.
Diplomarbeit / Masterarbeit - Institut für Nutztierwissenschaften (NUWI), BOKU-Universität für Bodenkultur, pp 40. UB BOKU obvsg FullText

Datenquelle: ZID Abstracts
Abstract:
The previous discovery of a single nucleotide substitution in intron 3 of the insulin-like growth factor 2 (IGF2) gene in pigs revealed a novel mechanism for regulation of muscle growth. This mutation disrupts binding of a transcription factor, ZBED6. This domesticated DNA transposon is located within the first intron of ZC3H11A gene, which encodes a poorly characterized zinc finger binding protein. In this project, we attempted to characterize the expression patterns and biological function of ZC3H11A. We hypothesized that ZC3H11A may play a role in RNA splicing and muscle development. ZC3H11A was silenced in mouse myoblast cell lines using RNA interference. At 48 h post-transfection, total RNA was isolated and the expression levels of genes associated with myogenesis (MYOGENIN, PAX7 and SRF) and IGF2 regulation (ZBED6 and BAHD1) were quantified by real-time PCR. Effects of gene silencing on cell proliferation and differentiation were also assessed. The alternative splicing of various genes was evaluated in ZC3H11A-silenced cells. Whole mount mouse embryo and placenta samples were prepared for immunohistochemistry (IHC) at embryonic days 7.5, 8.5, 10.5, 12.5 and 13.5 to visualize the expression pattern of ZC3H11A during mouse embryo development. Results showed that silencing ZC3H11A was associated with a down-regulation of MYOGENIN, PAX7 and BAHD1 mRNA (p < 0.05). As silencing of ZC3H11A targets the transcript containing ZBED6, ZBED6 mRNA was also down-regulated (p < 0.001). When ZBED6 was knocked down in C2C12 cells using siRNA, IGF2 mRNA was up-regulated (P<0.001) more than 2-fold. No aberrant RNA splicing was detected in ZC3H11A-silenced cells. Immunohistochemical staining revealed a nuclear localization for the ZC3H11A protein. The ZC3H11A protein was restricted to certain tissues during embryo development, particularly cartilage and muscle. ZC3H11A appears to be a factor important for mammalian development. Because of the integration of ZBED6 into the first intron of the ZC3H11A gene and use of a common promoter for transcription, it is difficult to elucidate the underlying mechanisms regarding regulation and function of these two unique genes.

Beurteilende(r): Sölkner Johann
1.Mitwirkender: Meszaros Gabor

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