Detection of Nucleic Acid-Nuclear Hormone Receptor Complexes with Mass Spectrometry

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Author(s) Bich, Claudia, Bovet, Cédric, Rochel, Natacha, Peluso-Iltis, Carole, Panagiotidis, Andreas, Nazabal, Alexis, Moras, Dino, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Detection of Nucleic Acid-Nuclear Hormone Receptor Complexes with Mass Spectrometry
Author(s) Bich, Claudia
Bovet, Cédric
Rochel, Natacha
Peluso-Iltis, Carole
Panagiotidis, Andreas
Nazabal, Alexis
Moras, Dino
Zenobi, Renato
Journal or Series Title Journal of the American Society for Mass Spectrometry
Volume Number 21
Issue Number 4
Start Page 635
End Page 645
ISSN 1044-0305
Publisher Elsevier Science
Publication Place New York
Publication Date 2010
Abstract Nuclear receptors, such as the retinoic acid receptor (RAR) or the 9-cis retinoic acid receptor (RXR), interact not only with their ligands but also with other types of receptors and with DNA. Here, two complementary mass spectrometry (MS) methods were used to study the interactions between retinoic receptors (RXR/RAR) and DNA: non-denaturing nano-electrospray (nanoESI MS), and high-mass matrix-assisted laser desorption ionization (MALDI MS) combined with chemical cross-linking. The RAR.RXR heterodimer was studied in the presence of a specific DNA sequence (DR5), and a specific RAR.RXR.DNA complex was detected with both MS techniques. RAR by itself showed no significant homodimerization. A complex between RAR and the double stranded DR5 was detected with nanoESI. After cross-linking, high-mass MALDI mass spectra showed that the RAR binds the single stranded DR5, and the RAR dimer binds both single and double stranded DR5. Moreover, the MALDI mass spectrum shows a larger RAR dimer signal in the presence of DNA. These results suggest that a gene-regulatory site on DNA can induce quaternary structural changes in a transcription factor such as RAR.
DOI 10.1016/j.jasms.2009.12.004
Additional Notes Received 9 August 2009, Revised 9 December 2009, Accepted 12 December 2009, Available online 28 December 2009
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
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NEBIS System Number 000541631
Source Database ID WOS-000276788700016
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@article{Bch2010,
  author = "Bich, Claudia and Bovet, C{\'{e}}dric and Rochel, Natacha and Peluso-Iltis, Carole and Panagiotidis, Andreas and Nazabal, Alexis and Moras, Dino and Zenobi, Renato",
  title = "{D}etection of {N}ucleic {A}cid-{N}uclear {H}ormone {R}eceptor {C}omplexes with {M}ass {S}pectrometry",
  journal = "Journal of the American Society for Mass Spectrometry",
  year = 2010,
  volume = "21",
  number = "4",
  pages = "635--645",
}


E-Citations record created: Mon, 03 May 2010, 07:23:00 CET