DNA-induced increase in the alpha-helical content of C/EBP and GCN4.
Leucine zipper proteins comprise a recently identified class of DNA binding proteins that contain a bipartite structural motif consisting of a "leucine zipper" dimerization domain and a segment rich in basic residues responsible for DNA interaction. Protein fragments encompassing the zipper plus basic region domains (bZip) have previously been used to determine the conformational and dynamic properties of this motif. In the absence of DNA, the coiled-coil portion is alpha-helical and dimeric, whereas the basic region is flexible and partially disordered. Addition of DNA containing a specific recognition sequence induces a fully helical conformation in the basic regions of these fragments. However, the question remained whether the same conformational change would be observed in native bZip proteins where the basic regions might be stabilized in an alpha-helical conformation even in the absence of DNA, through interactions with portions of the protein not included in the bZip motif. We have now examined the DNA-induced conformational transition for an intact bZip protein, GCN4, and for the bZip fragment of C/EBP with two enhancers that are differentially symmetric. Our results are consistent with the induced helical fork model wherein the basic regions are largely flexible in the absence of DNA and become fully helical in the presence of the specific DNA recognition sequence.
O'Neil
K T
KT
Central Research and Development Department, DuPont Company, Wilmington, Delaware 19880-0328.
Shuman
J D
JD
Ampe
C
C
DeGrado
W F
WF
D001483:Base Sequence; D022762:CCAAT-Enhancer-Binding Proteins; D002942:Circular Dichroism; D004247:DNA / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000187:drug effects; D005656:Fungal Proteins / Q000737:chemistry* / Q000187:drug effects; D016350:Leucine Zippers / Q000187:drug effects; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry* / Q000187:drug effects; D011487:Protein Conformation / Q000187:drug effects; D011494:Protein Kinases; D029701:Saccharomyces cerevisiae Proteins; D013329:Structure-Activity Relationship; D013431:Sulfates; D014157:Transcription Factors / Q000737:chemistry* / Q000187:drug effects
D022762:CCAAT-Enhancer-Binding Proteins; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D013431:Sulfates; D014157:Transcription Factors; D004247:DNA; D011494:Protein Kinases