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An investigation into the RNA-binding protein UNR and its interactors

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Ó Catnaigh, Pól (2016) An investigation into the RNA-binding protein UNR and its interactors. PhD thesis, University of Warwick.

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Official URL: http://webcat.warwick.ac.uk/record=b3093791~S15

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Abstract

Recent work linked the RNA-binding protein UNR to a number of human pathologies although little is currently known about how UNR functions in human cells. This thesis aims to elucidate how UNR functions by, among other things, discovering novel UNR-interacting proteins and transcripts and proteins that are differentially expressed in the presence or absence of UNR.

It is shown that UNR levels decrease with increasing cell confluency in cultured HeLa cells but that they increase with increasing confluency in the wild type TP53-containing U2OS cell line. UNR is shown to colocalise to stress granules with TP53 in arsenite-stressed HeLa cells.

A number of novel UNR-interacting proteins were discovered in three cell lines (HeLa, U2OS and SaOS-2), including HUWE1, NARR, SQSTM1 and LDB1. GO-term overrepresentation analysis confirmed that UNR is an RNA-binding protein as ‘RNA binding’ and ‘poly(A) RNA binding’ were the top two overrepresented molecular function GO terms by p-value across each of the cell types. Less expected overrepresented GO terms pertained to selenium metabolism and the extracellular exosome.

There was no evidence for conservation of UNR-interacting transcripts across the cell types but there were some similar significantly overrepresented GO terms among the respective UNR-interacting transcripts. These included terms pertaining to RNA and the nucleus. The most significant UNR-interacting transcript in HeLa cells was PABPC1 and that the PABP protein was also significantly upregulated following UNR knockdown in HeLa cells.

‘Poly (A) RNA binding’ was a significantly overrepresented GO term among proteins differentially regulated following UNR knockdown in HeLa and U2OS cells. ‘Adherens junction’ was another significantly overrepresented GO term using proteins that were higher in abundance in siUNR-treated HeLa cells and either higher or lower in abundance in either arsenite-stressed or unstressed U2OS cells. UNR was observed at cell-cell junctions in HeLa and U2OS cells by immunofluorescence microscopy.

Item Type: Thesis (PhD)
Subjects: Q Science > QP Physiology
Library of Congress Subject Headings (LCSH): RNA-protein interactions, Protein binding, Systems biology
Official Date: December 2016
Dates:
DateEvent
December 2016Submitted
Institution: University of Warwick
Theses Department: Systems Biology Doctoral Training Centre
Thesis Type: PhD
Publication Status: Unpublished
Supervisor(s)/Advisor: Anderson, Emma C. ; Ott, Sascha
Format of File: pdf
Extent: 1 volume (various pagings) : illustrations, charts
Language: eng

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