Complex formation of nuclear proteins with the RNA polymerase I promoter and repeated elements in the external transcribed spacer of Cucumis sativus ribosomal DNA.
Complex repetitive structures are located downstream of the transcription initiation site in the intergenic spacer (IGS) of the rRNA genes in Cucumis sativus (cucumber). In order to show that these repetitive elements of the 5'external transcribed spacer (ETS) are probably involved in transcriptional regulation as protein binding sites DNA-protein binding assays were carried out. The same proteins that recognize two binding sites in the promoter region analysed (upstream binding element between -164 and -105, and core promoter between -41 and +16) show binding affinity to the complex structures of the 5'external transcribed spacer. These proteins also seem to interact with the single strands of the respective DNA regions suggesting an effect on transcriptional regulation while the DNA is transcribed and, therefore, is single-stranded. Three proteins were isolated by affinity column chromatography; these proteins turned out to be much smaller (16, 22, and 24 kDa, respectively) than promoter and enhancer binding proteins in animal systems. Additionally, a 70-kDa protein could be characterized cooperating with a small segment of the repeated elements but not with the promoter.
Zentgraf
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U
Lehrstuhl für Allgemeine Genetik, Biologisches Institut, Tübingen, Germany.
Hemleben
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V
D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002846:Chromatography, Affinity; D016384:Consensus Sequence / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000235:genetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D012318:RNA Polymerase I / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid / Q000502:physiology*; D014158:Transcription, Genetic / Q000235:genetics
D004275:DNA, Ribosomal; D004268:DNA-Binding Proteins; D046911:Macromolecular Substances; D012318:RNA Polymerase I