This seminar will present a new rapid and robust method for single molecule analysis of DNA binding proteins from nuclear extracts (SMADNE) of human cells expressing a fluorescently tagged protein of interest (Schaich et al., Nucleic Acids Res. 2023 Apr 24;51(7):e39). SMADNE when combined with the LUMICKS C-trap provides unprecedented observations of DNA repair protein dynamics on damaged DNA. Over the last 18 months our group has been able to analyze the dwell times of 30 proteins or protein variants on DNA substrates using this approach. After a brief introduction to DNA damage and base excision repair, this seminar will discuss new unpublished data, including the DNA binding dynamics of PARP1 and PARP1 variants or DNA LIG3-XRCC1 on nicks in naked DNA and in a nucleosome. An important glycosylase, TDG, involved in active oxidative demethylation of 5mC through base excision repair will also be discussed. Finally, the non-specific DNA interactions and the binding behavior to 8-oxoG of purified eGFP-OGG1, purified eGFP-OGG1 added to extracts, and eGFP-OGG1 from human cell nuclear extracts will be directly compared.
A new single molecule approach to study DNA repair protein dynamics: seeing is believing Genetic Instability and Cancer
ORGANIZED BY IGH
International Summer-School on Epigenetics from 15/07/2024 until 19/07/2024 Genopolys, Montpellier - France
Meiosis and recombinationMBHD Dpt. Seminar Series
IGH Seminar Series of the Department “Molecular basis of Human Diseases” from 02/04/2024 until 11/10/2024 11:30 - 14:00 Genopolys amphitheater
GCB&D Dpt. Seminar Series
IGH Seminar Series of the Department “Genetics, Cell Biology and Development” from 19/02/2024 until 15/11/2024 Genopolys amphitheater
Seminars
Selective autophagy of DNA lesions promotes DNA repair and cell survival
11/10/2024 11:30 Genopolys AmphitheatreIchthyosporeans and the evolution of animal embryogenesis
15/11/2024 11:30 Genopolys amphitheaterCongresses
Transcription and Genome maintenance
09/09/2024 until 13/09/2024 Roscoff, France2024 RNase H meeting
16/09/2024 until 18/09/2024 Engelthal Ingelheim, GermanyCBS2 day 2024
26/09/2024 Université Montpellier, FrTweet