Department Molecular Genetics
Principal investigator Jurgen Marteijn
E-mail address j.marteijn@erasmusmc.nl
Website https://www.genomestability.nl/
How transcription-blocking DNA damage is recognized by CSB
Supervisor: Daria Repkina, d.repkina@erasmusmc.nl
This project investigates how cells protect their DNA, using advanced single-molecule tracking and confocal imaging to explore transcription-coupled DNA repair. Disruptions in transcription due to DNA damage can lead to cellular aging, apoptosis, or rare genetic diseases when DNA repair pathways fail. Cells counteract these threats with transcription-coupled nucleotide-excision repair (TC-NER), which clears transcription-blocking lesions to support cell health. This study focuses on CSB, a key regulatory protein in TC-NER, examining how it identifies and responds to DNA damage at a molecular level. Results show that CSB immobilizes at damage sites in direct response to DNA stress. This research offers a valuable opportunity for students to gain experience with state-of-the-art microscopy techniques in molecular biology.
Techniques
- Single-molecule tracking
- Quantitative analysis of molecular mobility
- Fixed and live-cell confocal imaging
- Cell culture
- Cytotoxicity evaluation
- Genome engineering (CRISPR-Cas9)
Further reading
https://www.nature.com/articles/s41580-019-0169-4
New regulators of Transcription-Coupled repair
Supervisor: Jurgen Marteijn, J.Marteijn@erasmusmc.nl
Unperturbed transcription of eukaryotic genes by RNA polymerase II (Pol II) is crucial for proper cell function and tissue homeostasis. However, the DNA template of Pol II is continuously challenged by DNA damage that can impede or block transcription. These transcription-blocking DNA damages can cause cellular dysfunction, senescence and apoptosis, eventually resulting in DNA damage-induced aging. Cells counteract these deleterious effects by transcription-coupled repair (TCR), which specifically removes DNA damage in our genes thereby safeguarding transcription. Our lab made important contributions in the TCR-field by identifying several new repair factors and uncovering their molecular mechanism, including UVSSA, ELOF1, HLTF and STK19. In this project we will study the involvement and the mode-of-action of new factors in TCR, that we identified in CRISPR/Cas9 or proteomics screens.
Techniques
- Cell culture
- CRISPR/Cas9 genome editing
- Molecular biology experiments
- Western blot
- Immunoprecipitation
- Immunofluorescence based repair assays
- Live cell imaging