Alternative splicing regulation ofdrosophila down syndrome celladhesion molecule gene
Başlık çevirisi mevcut değil.
- Tez No: 758606
- Danışmanlar: Belirtilmemiş.
- Tez Türü: Doktora
- Konular: Çevre Mühendisliği, Environmental Engineering
- Anahtar Kelimeler: Belirtilmemiş.
- Yıl: 2015
- Dil: İngilizce
- Üniversite: The University of Birmingham
- Enstitü: Yurtdışı Enstitü
- Ana Bilim Dalı: Belirtilmemiş.
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: 170
Özet
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Özet (Çeviri)
Alternative splicing of pre-mRNA is a prominent mechanism to increase protein diversity. The Drosophila homolog of human Down Syndrome Cell Adhesion Molecule (Dscam) encodes a cell surface protein of the immunoglobulin superfamily and represents an extraordinary example of alternative splicing. The Dscam gene comprises 95 alternatively spliced exons that are organized into 4 clusters, namely 4, 6, 9 and 17 which contain 12, 48, 33 and 2 variables respectively. Thus, the Dscam gene can potentially generate 38,016 different proteins. Dscam is functionally required for neuronal wiring in the nervous system, but also for phagocytosis of invading pathogens in the immune system. Currently, it is not understood how exons are prevented from being spliced together and how a specific exon is chosen. To understand how Dscam splicing is regulated, cis-acting elements and splicing regulators were analyzed. To make such analysis possible, reporter transgenes that recapitulate endogenous splicing in flies were characterized. Using a phylogenetic approach, inclusion levels of two different Drosophila species were compared: D. melanogster and D. virilis were analyzed for the similarity of splice sites to consensus splice sites in the variable exon 4 cluster correlates with inclusion levels. This data showed no correlation between splice site strength and inclusion levels. Since sequences in exon 4 cluster introns are not conserved, this suggests that exonic sequences in the variable exon 4 cluster contain the regulatory elements required for inclusion. Indeed, it has been confirmed by exchanging exon 4 with exon 9 variables utilising reporter transgenes. SR (serine/arginine-rich) and hnRNPs (heterogeneous nuclear ribonucleoproteins) proteins are important regulators of splicing. Since current models propose that variable clusters are kept in a repressed state and prevent the splicing together of adjacent exons; Dscam splicing was analyzed in loss and gain of SR and hnRNP protein function mutants. For both SR and hnRNP proteins, there was no significant impact on Dscam exon 9 splicing from manipulating individual genes. In bees, both Dscam and elav are alternatively spliced and have roles in neuronal wiring, raising the possibility that they are required for establishing new connections during memory formation. To test this hypothesis, bees, the preferred model organism, were tested to examine learning and memory. Here, the splicing pattern of exon variables during development, learning and memory were characterised to demonstrate how splicing patterns change. Overall, these findings demonstrate that exonic sequences play an important role in Dscam splicing regulation. Moreover, mutations in individual SR and hnRNP proteins demonstrated minimal impact on Dscam splicing. This suggests that these proteins act redundantly. In addition, alternatively spliced variables of Dscam and elav was first demonstrated in bees and their splicing changes were analyzed during learning and memory.
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