Identification of FUS RNA targets and the role of FUS-dependent RNA metabolism in ALS and cancer
| dc.contributor.author | Zhou, Yueqin | |
| dc.contributor.examiningcommittee | Davie, James (Biochemistry and Medical Genetics) Wilkins, John (Biochemistry and Medical Genetics) Marshall, Aaron (Biochemistry and Medical Genetics) McKenna, Sean (Chemistry) Bonham, Keith (University of Saskatchewan) | en_US |
| dc.contributor.supervisor | Hicks, Geoff (Biochemistry and Medical Genetics) | en_US |
| dc.date.accessioned | 2014-08-25T20:32:14Z | |
| dc.date.available | 2014-08-25T20:32:14Z | |
| dc.date.issued | 2013-10-31 | en_US |
| dc.date.issued | 2014-06-12 | en_US |
| dc.degree.discipline | Biochemistry and Medical Genetics | en_US |
| dc.degree.level | Doctor of Philosophy (Ph.D.) | en_US |
| dc.description.abstract | FUS/TLS is a multifunctional RNA binding protein. The RNA binding function of FUS is either lost in cancer associated FUS translocations or altered in amyotrophic lateral sclerosis (ALS)-associated FUS mutants, suggesting dysregulated FUS-dependent RNA metabolism may play central roles in disease pathogenesis. The goal of my thesis work was to elucidate the function of FUS in RNA metabolism, the underlying mechanisms, and its relevance to ALS and cancer. We identified FUS RNA targets on a genome wide scale in HeLa cells using CLIP-seq. Our data indicate that FUS targets RNA of genes encoding transcription, RNA metabolic processes, and genes associated with cancer and neurogenesis. Analysis of FUS RNA targets revealed that FUS regulates alternative splicing of cassette exons, processing of long intron containing transcripts, and ncRNA biogenesis. We specifically focused on the function of FUS in pre-mRNA cassette exon splicing and miRNA biogenesis. We discovered a novel FUS autoregulatory mechanism through the alternative splicing of its own exon 7 and nonsense mediated decay, which controls FUS protein homeostasis. We show ALS-FUS mutants are deficient in alternative splicing and autoregulation, which forms a feed-forward loop to exacerbate the pathological FUS accumulation in ALS. We demonstrated FUS regulates the expression of the oncogenic miRNA cluster miR-17-92. FUS interacts with the pri-miR-17-92 transcripts in vivo and appears to be associated with the promoter and the coding regions of miR-17-92 chromatin, suggesting a role of FUS in the cotranscriptional processing of miR-17-92. Moreover, we identified for the first time FUS-dependent miRNA-mRNA regulatory networks involving DNA damage repair, cell proliferation and cell cycle, suggesting dysregulated miRNA biogenesis may contribute to FUS-associated cancer. In conclusion, we identified FUS RNA targets on a genome-wide scale, revealed the function of FUS specifically in the alternative splicing of its own pre-mRNA and the biogenesis of miR-17-92. This work opens up new research avenues and enhances our understanding of FUS-dependent RNA metabolism. It provides experimental evidence demonstrating that dysregulated RNA processing may play central roles in FUS-associated cancer and ALS pathogenesis. The better understanding of FUS-dependent RNA metabolism also facilitates the development of novel therapeutic strategies to treat ALS and cancer. | en_US |
| dc.description.note | October 2014 | en_US |
| dc.identifier.citation | Zhou Y, Liu S, Liu G, Öztürk A, Hicks GG. ALS-Associated FUS Mutations Result in Compromised FUS Alternative Splicing and Autoregulation. PLoS Genet 2013; 9(10): e1003895. | en_US |
| dc.identifier.citation | Zhou Y, Liu S, Öztürk A, Hicks GG. FUS-regulated RNA Metabolism and DNA Damage Repair: Implications for Amyotrophic Lateral Sclerosis and Frontotemporal Dementia Pathogenesis. Rare Diseases 2014; 2:e29515. | en_US |
| dc.identifier.uri | http://hdl.handle.net/1993/23879 | |
| dc.language.iso | eng | en_US |
| dc.publisher | PLoS Genetics | en_US |
| dc.publisher | Rare Diseases | en_US |
| dc.rights | open access | en_US |
| dc.subject | FUS/TLS | en_US |
| dc.subject | RNA metabolism | en_US |
| dc.subject | alternative splicing | en_US |
| dc.subject | miRNA | en_US |
| dc.subject | amyotrophic lateral sclerosis | en_US |
| dc.subject | Cancer | en_US |
| dc.subject | ALS | |
| dc.title | Identification of FUS RNA targets and the role of FUS-dependent RNA metabolism in ALS and cancer | en_US |
| dc.type | doctoral thesis | en_US |