Understanding the role of RNA nucleases in human disease
<p>Congenital dyserythropoietic anaemia type-I (CDA-I) is a rare inherited disorder of erythropoiesis, in which patient erythroblasts uniquely display a pathognomonic pattern of heterochromatin, referred to as ‘spongy’ heterochromatin. The molecular basis of CDA-I remains unknown. Most cases o...
| Main Author: | |
|---|---|
| Other Authors: | |
| Format: | Thesis |
| Language: | English |
| Published: |
2021
|
| _version_ | 1817931817283485696 |
|---|---|
| author | Brolih, S |
| author2 | McHugh, P |
| author_facet | McHugh, P Brolih, S |
| author_sort | Brolih, S |
| collection | OXFORD |
| description | <p>Congenital dyserythropoietic anaemia type-I (CDA-I) is a rare inherited disorder of erythropoiesis, in which patient erythroblasts uniquely display a pathognomonic pattern of heterochromatin, referred to as ‘spongy’ heterochromatin. The molecular basis of CDA-I remains unknown. Most cases of CDA-I are caused by mutations in a protein of unknown function, termed Codanin-1. However, the recent identification of a second disease gene in CDA-I, namely C15orf41, has marked a breakthrough in providing new insights into CDA-I. This study describes the previously-uncharacterised protein C15orf41 as a novel RNA exonuclease that associates in vivo and in vitro with Codanin-1. Using the newly generated CDA-I disease model in an erythroid-specific HUDEP-2 cell background, C15orf41 and Codanin-1 complex formation is determined at physiologically relevant levels. This interaction is then also confirmed in vitro. The C15orf41-Codanin-1 complex is a versatile RNA nuclease complex, able to digest a variety of RNA structures. We also show the association and nucleolytic activity of C15orf41-Codanin-1 are disrupted by the introduction of CDA-I patient mutations p. Y94C and p. L178Q in C15orf41. This suggests a direct link between the disease phenotype observed and C15orf41’s novel RNAse activity, a potential new target for alternative treatment development.</p>
<p>In light of the 2020 SARS-CoV-2 global pandemic, the focus of this study was broadened to perform essential research on the SARS-CoV-2 virus, in particular the nsp14-nsp10 RNA nuclease complex. This work provides the first complete biochemical activity report for the SARS-CoV-2 nsp14-nsp10 complex, characterising its exonuclease activity as well as a identifying a novel, endonuclease activity. These results have served as a basis for inhibitor development and screening, essential considering the global health impact of this virus.</p> |
| first_indexed | 2024-03-07T07:11:43Z |
| format | Thesis |
| id | oxford-uuid:0911808e-063e-4b99-818e-83e15be6f8d0 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-12-09T03:28:03Z |
| publishDate | 2021 |
| record_format | dspace |
| spelling | oxford-uuid:0911808e-063e-4b99-818e-83e15be6f8d02024-12-01T10:58:29ZUnderstanding the role of RNA nucleases in human diseaseThesishttp://purl.org/coar/resource_type/c_db06uuid:0911808e-063e-4b99-818e-83e15be6f8d0EnglishHyrax Deposit2021Brolih, SMcHugh, PBabbs, C<p>Congenital dyserythropoietic anaemia type-I (CDA-I) is a rare inherited disorder of erythropoiesis, in which patient erythroblasts uniquely display a pathognomonic pattern of heterochromatin, referred to as ‘spongy’ heterochromatin. The molecular basis of CDA-I remains unknown. Most cases of CDA-I are caused by mutations in a protein of unknown function, termed Codanin-1. However, the recent identification of a second disease gene in CDA-I, namely C15orf41, has marked a breakthrough in providing new insights into CDA-I. This study describes the previously-uncharacterised protein C15orf41 as a novel RNA exonuclease that associates in vivo and in vitro with Codanin-1. Using the newly generated CDA-I disease model in an erythroid-specific HUDEP-2 cell background, C15orf41 and Codanin-1 complex formation is determined at physiologically relevant levels. This interaction is then also confirmed in vitro. The C15orf41-Codanin-1 complex is a versatile RNA nuclease complex, able to digest a variety of RNA structures. We also show the association and nucleolytic activity of C15orf41-Codanin-1 are disrupted by the introduction of CDA-I patient mutations p. Y94C and p. L178Q in C15orf41. This suggests a direct link between the disease phenotype observed and C15orf41’s novel RNAse activity, a potential new target for alternative treatment development.</p> <p>In light of the 2020 SARS-CoV-2 global pandemic, the focus of this study was broadened to perform essential research on the SARS-CoV-2 virus, in particular the nsp14-nsp10 RNA nuclease complex. This work provides the first complete biochemical activity report for the SARS-CoV-2 nsp14-nsp10 complex, characterising its exonuclease activity as well as a identifying a novel, endonuclease activity. These results have served as a basis for inhibitor development and screening, essential considering the global health impact of this virus.</p> |
| spellingShingle | Brolih, S Understanding the role of RNA nucleases in human disease |
| title | Understanding the role of RNA nucleases in human disease |
| title_full | Understanding the role of RNA nucleases in human disease |
| title_fullStr | Understanding the role of RNA nucleases in human disease |
| title_full_unstemmed | Understanding the role of RNA nucleases in human disease |
| title_short | Understanding the role of RNA nucleases in human disease |
| title_sort | understanding the role of rna nucleases in human disease |
| work_keys_str_mv | AT brolihs understandingtheroleofrnanucleasesinhumandisease |