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Journal Article

Atomic-resolution mapping of transcription factor-DNA interactions by femtosecond laser crosslinking and mass spectrometry

MPS-Authors
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Reim,  Alexander
Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society;

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Mann,  Matthias
Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society;

/persons/resource/persons195388

Wierer,  Michael
Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society;

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Fulltext (public)

s41467-020-16837-x.pdf
(Publisher version), 2MB

Supplementary Material (public)

41467_2020_16837_MOESM1_ESM.pdf
(Supplementary material), 7MB

41467_2020_16837_MOESM2_ESM.pdf
(Supplementary material), 843KB

41467_2020_16837_MOESM3_ESM.docx
(Supplementary material), 25KB

41467_2020_16837_MOESM4_ESM.pdf
(Supplementary material), 3MB

41467_2020_16837_MOESM5_ESM.xlsx
(Supplementary material), 168KB

41467_2020_16837_MOESM6_ESM.pdf
(Supplementary material), 2MB

Citation

Reim, A., Ackermann, R., Font-Mateu, J., Kammel, R., Beato, M., Nolte, S., et al. (2020). Atomic-resolution mapping of transcription factor-DNA interactions by femtosecond laser crosslinking and mass spectrometry. Nature Communications, 11(1): 3019. doi:10.1038/s41467-020-16837-x.


Cite as: http://hdl.handle.net/21.11116/0000-0007-18AD-C
Abstract
Transcription factors (TFs) regulate target genes by specific interactions with DNA sequences. Detecting and understanding these interactions at the molecular level is of fundamental importance in biological and clinical contexts. Crosslinking mass spectrometry is a powerful tool to assist the structure prediction of protein complexes but has been limited to the study of protein-protein and protein-RNA interactions. Here, we present a femtosecond laser-induced crosslinking mass spectrometry (fliX-MS) workflow, which allows the mapping of protein-DNA contacts at single nucleotide and up to single amino acid resolution. Applied to recombinant histone octamers, NF1, and TBP in complex with DNA, our method is highly specific for the mapping of DNA binding domains. Identified crosslinks are in close agreement with previous biochemical data on DNA binding and mostly fit known complex structures. Applying fliX-MS to cells identifies several bona fide crosslinks on DNA binding domains, paving the way for future large scale ex vivo experiments.