UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

RNA target recognition by IMP1 – a molecular investigation at the transcriptome level

Gallagher, Christopher; (2018) RNA target recognition by IMP1 – a molecular investigation at the transcriptome level. Doctoral thesis (Ph.D), UCL (University College London).

[img] Text
Christopher_Gallagher_Corrected_Thesis.pdf - Accepted version
Access restricted to UCL open access staff until 1 September 2021.

Download (10MB)

Abstract

RNA binding proteins orchestrate the assembly of macromolecular RNA-protein complexes that regulate post-transcriptional regulation. Disruption of these finely tuned processes can result in an array of diseases. RNA binding proteins are multidomain proteins organised into a modular structure. It is through this structure that they can recognise a vast repertoire of RNA targets by employing combinatorial binding. However, this mechanism is poorly understood. The IMP1 protein provides a model system in which we can investigate how domain sequence specificity and combinatorial binding define in vivo RNA selection. IMP1 recognises RNA via six putative RNA binding domains (two N-terminal RNA recognition motifs and four C-terminal K-homology domains). To date, IMP1 is known to bind a diverse range of RNA targets, both in homeostatic cellular events and in cancer. However, detailed information as to how IMP1 recognises these targets, especially at the individual domain level in context of the full-length protein is lacking. I have implemented a structural driven mutational approach to modify the RNA recognition properties of the individual RNA binding domains of the IMP1 protein. I have successfully generated iCLIP libraries for mutant IMP1 proteins where each KH domain was mutated in turn to inhibit RNA binding. From comparative analysis of these data sets I have begun to explore in vivo RNA target selection at the individual domain level and observed an altered RNA binding pattern to the ACTB mRNA. I have investigated in vitro, the RNA sequence specificity of the KH3-KH4 and the RRM1-RRM2 domains. This has led to the design of a mutant which shifts the RNA specificity of the KH3 domain. In addition to the identification that the RRM domains of IMP1 and IMP3 specifically recognise different RNA target sequences with different affinities, and the key residues involved in RNA recognition. These findings will aid in the further understanding of IMP in vivo RNA target selection.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: RNA target recognition by IMP1 – a molecular investigation at the transcriptome level
Event: UCL (University College London)
Language: English
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
URI: https://discovery.ucl.ac.uk/id/eprint/10054254
Downloads since deposit
2Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item