GAGA factor binding to DNA via a single trinucleotide sequence element.
GAGA transcription factor (GAF) is an essential protein in Drosophila , important for the transcriptional regulation of numerous genes. GAF binds to GA repeats in the promoters of these genes via a DNA-binding domain containing a single zinc finger. While GAF binding sites are typically composed of 3.5 GA repeats, the Drosophila hsp70 gene contains much smaller elements, some of which are as little as three bases (GAG) in length. Interestingly, the binding of GAF to more distant trinucleotide elements is relatively strong and not appreciably affected by the removal of larger GA arrays in the promoter. Moreover, a simple synthetic GAG sequence is sufficient to bind GAF in vitro . Here we directly compare the affinity of GAF for different sequence elements by immunoprecipitation and gel mobility shift analysis. Furthermore, our measures of the concentration of GAF in vivo indicate that it is a highly abundant nuclear protein, prevalent enough to occupy a sizable fraction of correspondingly abundant trinucleotide sites.
Wilkins
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RC
Section of Genetics and Development, Cornell University, Ithaca, NY 14853, USA.
Lis
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JT
D000818:Animals; D001665:Binding Sites; D002467:Cell Nucleus; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D004331:Drosophila melanogaster; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D018398:Homeodomain Proteins / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000378:metabolism*
D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D018840:HSP70 Heat-Shock Proteins; D018398:Homeodomain Proteins; D009711:Nucleotides; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; C063460:Trl protein, Drosophila; D004247:DNA