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Preliminary investigation of the interaction between the R2R3 DNA binding domain of the oncoprotein c‐Myb and DNA fragments
Author(s) -
Jamin Nadège,
Le Tilly Véronique,
Zargarian Loussinee,
Bostad Anne,
BesançonYoshpe Iris,
Lirsac PierreNoël,
Gabrielsen Odd S.,
Toma Flavio
Publication year - 1996
Publication title -
international journal of quantum chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.484
H-Index - 105
eISSN - 1097-461X
pISSN - 0020-7608
DOI - 10.1002/(sici)1097-461x(1996)59:4<333::aid-qua7>3.0.co;2-z
Subject(s) - dna , chemistry , myb , dissociation constant , base pair , titration , chemical shift , crystallography , fluorescence anisotropy , stereochemistry , nuclear magnetic resonance spectroscopy , fluorescence , dissociation (chemistry) , proton nmr , biochemistry , transcription factor , physics , gene , receptor , quantum mechanics , membrane
The interaction between the R2R3 DNA binding domain of the oncoprotein c‐Myb and oligodeoxynucleotides was investigated by 1 H‐ NMR spectroscopy and fluorescence anisotropy assays. Titration of 12 and 16 base‐pair DNA fragments containing the TAACGGTC sequence with R2R3 revealed the presence of two complexed forms (in a 40/60 ratio): either two complexes or two conformations in slow exchange at the NMR chemical shift time scale. The largest variations of imino proton chemical shifts were observed for the imino proton of the base pairs 2, 3, 4 and 6 of the DNA sequence, suggesting a direct involvement of these base pairs in the interaction. Using fluorescence anisotropy measurements, a dissociation constant of 5.12 ± 1.49 n M for the specific DNA‐R2R3 complex was found, whereas a value of 2.7 ± 0.1 μ M was determined for the nonspecific complex. © 1996 John Wiley & Sons, Inc.