PMID: 8948639Nov 15, 1996Paper

DNA binding of Jun and Fos bZip domains: homodimers and heterodimers induce a DNA conformational change in solution

Nucleic Acids Research
M JohnM Schnarr

Abstract

We constructed plasmids encoding the sequences for the bZip modules of c-Jun and c-Fos which could then be expressed as soluble proteins in Escherichia coli. The purified bZip modules were tested for their binding capacities of synthetic oligonucleotides containing either TRE or CRE recognition sites in electrophoretic mobility shift assays and circular dichroism (CD). Electrophoretic mobility shift assays showed that bZip Jun homodimers and bZip Jun/Fos heterodimers bind a collagenase-like TRE (CTGACTCAT) with dissociation constants of respectively 1.4 x 10(-7) M and 5 x 10(-8) M. As reported earlier [Patel et al. (1990) Nature 347, 572-575], DNA binding induces a marked change of the protein structure. However, we found that the DNA also undergoes a conformational change. This is most clearly seen with small oligonucleotides of 13 or 14 bp harboring respectively a TRE (TGACTCA) or a CRE (TGACGTCA) sequence. In this case, the positive DNA CD signal at 280 nm increases almost two-fold with a concomitant blue-shift of 3-4 nm. Within experimental error the same spectral changes are observed for TRE and CRE containing DNA fragments. The spectral changes observed with a non-specific DNA fragment are weaker and the signal of free DN...Continue Reading

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Citations

Feb 16, 2006·Nucleic Acids Research·Seiji TateyamaHiroshi Yanagawa
Oct 23, 2008·Archives of Biochemistry and Biophysics·Kenneth L SeldeenAmjad Farooq
Aug 23, 2008·Protein Expression and Purification·Natalie A CiaccioJennifer S Laurence
Sep 20, 2005·The Plant Journal : for Cell and Molecular Biology·Corinna ThurowChristiane Gatz
Mar 25, 2014·Biochimica Et Biophysica Acta·Steven Fletcher, Edward V Prochownik
Mar 5, 2008·Archives of Biochemistry and Biophysics·Kenneth L SeldeenAmjad Farooq
Jun 2, 2007·Current Biology : CB·Mikhail MartchenkoMalcolm Whiteway
Sep 10, 2013·Biophysical Chemistry·Kristin S GrußmayerDirk-Peter Herten
Jan 12, 2013·Biochemistry·Antonia T De Jong
Mar 7, 2021·International Journal of Molecular Sciences·Aikaterini KefalaMichael Kokkinidis

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