Interaction of TRF2-Rap1 with telomeric chromatin in the context of aging and cancer
Telomeric DNA form the ends of eukaryotic DNA. It is a tandem array of [TTAGGG]n, with a 3’ G-rich overhang. Telomeric nucleosomes form unique nucleoprotein structures with the shelterin complex consisting of 6 components; TRF1, TRF2, POT1, RAP1, TIN2, TPP1. Shelterin plays a crucial role in prevent...
| Main Author: | |
|---|---|
| Other Authors: | |
| Format: | Final Year Project (FYP) |
| Language: | English |
| Published: |
Nanyang Technological University
2024
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/178191 |
| _version_ | 1826127928443797504 |
|---|---|
| author | Woo, Jia Qi |
| author2 | Lars Nordenskiöld |
| author_facet | Lars Nordenskiöld Woo, Jia Qi |
| author_sort | Woo, Jia Qi |
| collection | NTU |
| description | Telomeric DNA form the ends of eukaryotic DNA. It is a tandem array of [TTAGGG]n, with a 3’ G-rich overhang. Telomeric nucleosomes form unique nucleoprotein structures with the shelterin complex consisting of 6 components; TRF1, TRF2, POT1, RAP1, TIN2, TPP1. Shelterin plays a crucial role in preventing inappropriate detection of telomeres as DNA breaks, and subsequent activation of damage repair pathways. Liquid-liquid phase separation induced by shelterin is proposed to be a mechanism underlying the dynamic behaviour of telomeres. Telomeres can switch between a protected and unprotected state to regulate specific access of factors to telomeres. Although the dynamic behaviour is integral to telomere integrity, little is understood about the behaviour.
Here, we characterise the dynamic behaviour of biomolecular condensates formed by telomeric nucleosomes under the influence of the TRF2-Rap1 complex. Full length hTRF2 and hRap1 were purified to derive the TRF-Rap1 complex. Nucleosome arrays were reconstituted with 10 x 157 base pairs of [TTAGGG]n and human histone octamers. Confocal microscopy was employed to study condensate behaviour. Condensates exhibit dynamicity up to TRF2-Rap1: Nucleosome = 2. Condensates formed with increasing TRF2-Rap1 display progressive rigidity. Our studies demonstrate that varying stoichiometries of TRF2-Rap1 complex at telomeric nucleosomes mediate condensate dynamicity. |
| first_indexed | 2024-10-01T07:16:32Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/178191 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:16:32Z |
| publishDate | 2024 |
| publisher | Nanyang Technological University |
| record_format | dspace |
| spelling | ntu-10356/1781912024-06-10T15:32:38Z Interaction of TRF2-Rap1 with telomeric chromatin in the context of aging and cancer Woo, Jia Qi Lars Nordenskiöld School of Biological Sciences LarsNor@ntu.edu.sg Medicine, Health and Life Sciences Telomeric DNA form the ends of eukaryotic DNA. It is a tandem array of [TTAGGG]n, with a 3’ G-rich overhang. Telomeric nucleosomes form unique nucleoprotein structures with the shelterin complex consisting of 6 components; TRF1, TRF2, POT1, RAP1, TIN2, TPP1. Shelterin plays a crucial role in preventing inappropriate detection of telomeres as DNA breaks, and subsequent activation of damage repair pathways. Liquid-liquid phase separation induced by shelterin is proposed to be a mechanism underlying the dynamic behaviour of telomeres. Telomeres can switch between a protected and unprotected state to regulate specific access of factors to telomeres. Although the dynamic behaviour is integral to telomere integrity, little is understood about the behaviour. Here, we characterise the dynamic behaviour of biomolecular condensates formed by telomeric nucleosomes under the influence of the TRF2-Rap1 complex. Full length hTRF2 and hRap1 were purified to derive the TRF-Rap1 complex. Nucleosome arrays were reconstituted with 10 x 157 base pairs of [TTAGGG]n and human histone octamers. Confocal microscopy was employed to study condensate behaviour. Condensates exhibit dynamicity up to TRF2-Rap1: Nucleosome = 2. Condensates formed with increasing TRF2-Rap1 display progressive rigidity. Our studies demonstrate that varying stoichiometries of TRF2-Rap1 complex at telomeric nucleosomes mediate condensate dynamicity. Bachelor's degree 2024-06-05T08:11:02Z 2024-06-05T08:11:02Z 2024 Final Year Project (FYP) Woo, J. Q. (2024). Interaction of TRF2-Rap1 with telomeric chromatin in the context of aging and cancer. Final Year Project (FYP), Nanyang Technological University, Singapore. https://hdl.handle.net/10356/178191 https://hdl.handle.net/10356/178191 en application/pdf Nanyang Technological University |
| spellingShingle | Medicine, Health and Life Sciences Woo, Jia Qi Interaction of TRF2-Rap1 with telomeric chromatin in the context of aging and cancer |
| title | Interaction of TRF2-Rap1 with telomeric chromatin in the context of aging and cancer |
| title_full | Interaction of TRF2-Rap1 with telomeric chromatin in the context of aging and cancer |
| title_fullStr | Interaction of TRF2-Rap1 with telomeric chromatin in the context of aging and cancer |
| title_full_unstemmed | Interaction of TRF2-Rap1 with telomeric chromatin in the context of aging and cancer |
| title_short | Interaction of TRF2-Rap1 with telomeric chromatin in the context of aging and cancer |
| title_sort | interaction of trf2 rap1 with telomeric chromatin in the context of aging and cancer |
| topic | Medicine, Health and Life Sciences |
| url | https://hdl.handle.net/10356/178191 |
| work_keys_str_mv | AT woojiaqi interactionoftrf2rap1withtelomericchromatininthecontextofagingandcancer |