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Title: Investigation of a novel Ercc1 transcript predominant in skin
Author: Simpson, Yvonne A.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2004
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In this project the skin-specific pattern of expression was studied by transfecting Ercc1 null cells with the minigenes. The Ercc1 null cells used were murine embryonic fibroblasts, PF24+#12D-4; Chinese hamster ovary cells, CHO43.3B (TG1); and murine keratinocytes; Ker. (-/-). IN survival studies were performed on transfected cells and RT-PCR was used to study the pattern of Ercc1 expression. Transfection with the Ercc1 minigenes corrected the IN sensitivity of the null cell lines. The Ercc1 expression pattern was usually, but not always; appropriate for the cell type transfected. Comparisons between the Ercc1 transcription of in vitro cultures and in vivo tissues taken from various stages of murine development were made by means of northern blotting. Irradiation of transfected cells and primary cultures with UV-B. UV-C and visible light was performed in an attempt to identify possible sources of transcript induction. Small changes in Ercc1 transcription following irradiation of primary cultures were observed, but these changes did not conclusively prove that visible light or UV could induce Ercc1 transcription. Serum starvation experiments were performed upon primary cultures to further study differences in Ercc1 transcription in vitro. Release from serum starvation resulted in increased skin-specific transcript production and total Ercc1 expression. The size of the skin-specific Ercc1 transcript in keratinocytes was found to be larger than that of fibroblasts. Sequencing demonstrated that the size difference was in the (CT)n repeat region of the gene. This region was deleted from one of the Ercc1 minigenes mentioned and results indicated that it may be required for correct expression of the novel transcript.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available