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Title: Functional analysis of novel protein, PIERCE1, in motile ciliogenesis
Author: Anujan, Priyanka
ISNI:       0000 0004 7960 368X
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2019
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Motile cilia are localised to tissues and cells where fluid movement and cellular locomotion is required. Mutations in genes associated with ciliogenesis and cilia motility give rise to diseases called ciliopathies. Primary Ciliary Dyskinesia (PCD), a heterogeneous genetic disorder, is the most common form of ciliopathy that arises from defects in motile cilia. Several systematic approaches have led to the identification of numerous genes with putative function in ciliogenesis and ciliary motility. We interrogated existing data and identified several novel candidate genes temporally associated with ciliogenesis. Expression of these genes were analysed in mouse airway epithelial cells during mucociliary differentiation at the air liquid interface (ALI) and different mouse tissues. This thesis focuses on a poorly characterized gene encoding the protein 'PIERCE1'. Transcriptional analysis of Pierce1 revealed an expression pattern temporally associated with ciliogenesis during differentiation of ALI mouse airway epithelial cells. Pierce1 also shows enriched expression in motile ciliated mouse tissues. Transient morpholino knock down of pierce1 in zebrafish showed phenotypes consistent with abnormalities in motile cilia and live imaging showed severe cilia motility defects in Kupffer's vesicle. Finally, we generated maternal zygotic loss-of-function alleles at the zebrafish pierce1 locus using the CRISPR/Cas9. These mutants showed mild laterality defects. A custom-made antibody against mouse full length PIERCE1 protein, was used to carry out immunofluorescence microscopy on ALI cultured mouse airway epithelial cells. It revealed that PIERCE1 is a cytoplasmic protein specifically expressed in motile ciliated cells. A yeast 2-hybrid assay carried out on human lung and testis libraries identified PIAS2, as a possible interacting partner of PIERCE1. With these findings, we propose that PIERCE1 may be involved in the assembly and transport of components required for cilia motility.
Supervisor: Bingle, Colin D. ; Bingle, Lynne ; Roy, Sudipto Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available