Xenbase Image ID: 132948

Figure 4 DNA/protein interactions. A,gel mobility shifts were performed with the bacterially expressed Xvent-2 homeodomain (HD) on the +1738/+1969 intron target sequence. 1, lane 1 contains free DNA;lanes 2–4 contain increasing amounts (6.25, 12.5, and 25 ng) of Xvent-2 HD. 2, addition of increasing amounts of unlabeled target sequence (specific inhibitor) leads to a loss of shift of the labeled target. 3, addition of an unspecific competitor (−69/+54 DNA fragment) does not prevent shifting.B, DNase I footprint of Xvent-2 HD was carried out for both strands on the +1738/+1969 intron 2 fragment. Lane 1 shows the A/G chemical sequencing reaction, lane 2 contains DNase I-digested free DNA. Increasing amounts (25, 75, and 225 ng) of Xvent-2 HD were added prior to DNase I digestion in lanes 3–5. Protected regions (indicated by lines) correspond for both strands and contain two copies of the motif 5′-CTAATT-3′ (underlined). Gene sequence is shown from nucleotide positions +1820 to + 1879. C, best fit alignment of theX. laevis Xvent-2-binding site with the human and mouseBMP-4 gene (number of matches: 13; allowed mismatches: 1). Note that a 13-bp match (including one T to C transition) is found within the proximal promoter of both the human (accession numberU43842) and the mouse BMP-4 genes (accession numberL47480). Image published in: Schuler-Metz A et al. (2000) Copyright © 2000. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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