Role of UCHL1 in regulating gene expression in prostate cancer cells
| dc.contributor.author | Ilic, Aleksandar | |
| dc.contributor.examiningcommittee | McManus, Kirk (Biochemistry and Medical Genetics) Rastegar, Mojgan (Biochemistry and Medical Genetics) Myal, Yvonne (Pathology) | en_US |
| dc.contributor.supervisor | Davie, Jim (Biochemistry and Medical Genetics) | en_US |
| dc.date.accessioned | 2014-08-28T16:58:54Z | |
| dc.date.available | 2014-08-28T16:58:54Z | |
| dc.date.issued | 2014-08-28 | |
| dc.degree.discipline | Biochemistry and Medical Genetics | en_US |
| dc.degree.level | Master of Science (M.Sc.) | en_US |
| dc.description.abstract | Ubiquitin C-terminal hydrolase L1 (UCHL1) is a multifunctional protein primarily expressed in neuronal cells and involved in numerous cellular processes. UCHL1 has been linked with neurodegenerative diseases and a wide range of cancers but its specific role remains unknown. Previous UCHL1 knockdown studies have shown that UCHL1 controls the expression of pro- and anti-apoptotic genes as well as genes involved in cell cycle regulation but it is unknown how UCHL1 regulates these genes. We have shown that UCHL1 is cross-linked to DNA in DU145 but not in LNCaP or PC3 prostate cancer cells. Therefore, we hypothesized that UCHL1 regulates the expression of pro- or anti-apoptotic genes as well as the genes involved in the cell cycle through its interaction with DNA. By utilizing ChIP and ChIP-seq analyses it is possible to determine the UCHL1 target sequences on the genomic DNA. It was shown that UCHL1 is only expressed in DU145 but not in LNCaP, PC3 or C4-2 prostate cancer cell lines. Additionally, UCHL1 is expressed and cross-linked to DNA in HEK293T cells. It is believed that UCHL1 is silenced by upstream promoter methylation when it is not expressed. However, treatment with the epigenetic drugs 5-aza-2′-deoxycytidine and trichostatin A (TSA) did not result in induction of UCHL1 expression in LNCaP, PC3 or C4-2 prostate cancer cell lines. UCHL1 is also associated with p53. However, ChIP assay results have shown that UCHL1 and p53 do not bind to genomic DNA of upstream promoter regions CDKN1A and BAX genes. Additionally, through UCHL1 ChIP-seq analyses in DU145 and HEK293T cells, we discovered that UCHL1 co-localizes to the DNA with the shelterin complex shedding light on a new role of UCHL1 that has never been described before. | en_US |
| dc.description.note | October 2014 | en_US |
| dc.identifier.uri | http://hdl.handle.net/1993/23912 | |
| dc.language.iso | eng | en_US |
| dc.rights | open access | en_US |
| dc.subject | UCHL1 | en_US |
| dc.subject | Prostate cancer | en_US |
| dc.subject | Next-generation sequencing | en_US |
| dc.subject | Chromatin immunoprecipitation | en_US |
| dc.subject | Epigenetic drugs | en_US |
| dc.title | Role of UCHL1 in regulating gene expression in prostate cancer cells | en_US |
| dc.type | master thesis | en_US |