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dc.contributor.authorMofatteh, Mohammad
dc.date.accessioned2018-05-21T09:45:53Z
dc.date.available2018-05-21T09:45:53Z
dc.date.issued2018-05-01
dc.date.submitted2018-03-31
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/275986
dc.description.abstractIn recent years it has become apparent that neuronal development and function relies not just on the regulation of transcription but also on post-transcriptional events. Two prevalent mRNA-based regulatory mechanisms in neurons are asymmetric mRNA localisation and the generation of different 3’UTR isoforms by alternative polyadenylation (APA). While experiments in mammalian systems indicate that subcellular mRNA localisation plays an important role in regulating local expression of proteins in neuronal processes, little is known about how mRNAs reach their destinations. It has been proposed that APA allows the production of mRNA isoforms with different roles. However, the importance of 3’UTR extensions has not been addressed in detail, particularly at the organismal level. In my PhD, I investigated the mechanisms of mRNA localisation and functional consequences of APA using the Drosophila embryonic nervous system as a genetically tractable model. I screened for mRNAs that localise in embryonic axons using an available transgenic library of 3’UTR sequences, as well as publically available in situ hybridisation data. I found that Ankyrin2 (Ank2) mRNA localises in Drosophila embryonic sensory neurons, and showed that this is dependent on the kinesin-1 motor and microtubules. These data reveal an active mRNA transport system in embryonic neurons. I also showed that the Ank2 mRNA has an extended 3’UTR that is found in axons, suggesting that APA could be relevant to axonal functions of Ank2. I demonstrated that while mRNA molecules could still localise to axons upon CRISPR-Cas9-mediated deletion of the Ank2 3’UTR extension, a fraction of the mutant embryos had a disrupted nervous system. Interestingly, embryos that lack the ability to make Ank2 protein have an overtly normal embryonic nervous system. This observation reveals that the extension does not simply promote Ank2 protein function. Further experiments revealed that the extended 3’UTR is required for efficient locomotion of adult flies. While the exact function of the Ank2 3’UTR extension requires future investigation, I show that it is unlikely to be associated with the trafficking of associated proteins into axons. RNA affinity purifications from embryonic extracts provide evidence that the 3’UTR extension selectively binds conserved RNA-binding proteins. I speculate that the extension plays a role in regulating axonal morphogenesis by regulating the relative expression level of different Ank2 protein isoforms.
dc.description.sponsorshipLMB Cambridge Scholarship
dc.language.isoen
dc.rightsAll rights reserved
dc.rightsAll Rights Reserveden
dc.rights.urihttps://www.rioxx.net/licenses/all-rights-reserved/en
dc.subjectDrosophila
dc.subjectmRNA
dc.subjectNeuron
dc.subjectposttranscriptiona modification
dc.subjectmRNA localisation
dc.subjectembryo
dc.subjectAnkyrin
dc.subjectAnk2
dc.subjectSensory neuron
dc.titleMechanism of mRNA localisation and posttranscriptional modification in Drosophila melanogaster embryonic neurons
dc.typeThesis
dc.type.qualificationlevelDoctoral
dc.type.qualificationnameDoctor of Philosophy (PhD)
dc.publisher.institutionUniversity of Cambridge
dc.publisher.departmentMRC Laboratory of Molecular Biology
dc.date.updated2018-05-20T17:20:04Z
dc.identifier.doi10.17863/CAM.23266
dc.contributor.orcidMofatteh, Mohammad [0000-0002-2458-0447]
dc.publisher.collegeWolfson
dc.type.qualificationtitleBiological Science at MRC-LMB
cam.supervisorBullock, Simon
cam.supervisorRoeper, Katja
cam.supervisorLandgraf, Matthias
cam.thesis.fundingtrue
rioxxterms.freetoread.startdate2019-05-21


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