Identification of two novel arginine binding DNAs.
RNA tertiary structure is known to play critical roles in RNA-protein recognition and RNA function. To examine how DNA tertiary structure might relate to RNA structure, we performed in vitro selection experiments to identify single-stranded DNAs that specifically bind arginine, and compared the results with analogous experiments performed with RNA. In the case of RNA, a motif related to the arginine binding site in human immunodeficiency virus TAR RNA was commonly found, whereas in the case of DNA, two novel motifs and no TAR-like structures were found. One DNA motif, found in approximately 40% of the cloned sequences, forms of hairpin structure with a highly conserved 10 nucleotide loop, whereas the second motif is especially rich in G residues. Chemical interference and mutagenesis experiments identified nucleotides in both motifs that form specific arginine binding sites, and dimethylsulfate footprinting experiments identified single guanine residues in both that are protected from methylation in the presence of arginine, suggesting possible sites of arginine contact or conformational changes in the DNAs. Circular dichroism experiments indicated that both DNAs undergo conformational changes upon arginine binding and that the arginine guanidinium group alone is responsible for binding. A model for the G-rich motif is proposed in which mixed guanine and adenine quartets may form a novel DNA structure. Arginine binding DNAs and RNAs should provide useful model systems for studying nucleic acid tertiary structure.
Harada
K
K
Department of Biochemistry and Biophysics, University of California at San Francisco 94143-0448, USA.
Frankel
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AD
D001120:Arginine / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D001665:Binding Sites; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D018983:DNA Footprinting; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000378:metabolism*; D015756:Genes, tat; D019791:Guanidine; D006146:Guanidines / Q000378:metabolism; D006678:HIV / Q000235:genetics; D016325:HIV Long Terminal Repeat / Q000235:genetics; D006801:Humans; D008745:Methylation; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D012313:RNA / Q000737:chemistry* / Q000378:metabolism*; D017421:Sequence Analysis; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism
D004277:DNA, Single-Stranded; D006146:Guanidines; D009841:Oligonucleotides; D012313:RNA; D001120:Arginine; D015719:Single-Strand Specific DNA and RNA Endonucleases; D019791:Guanidine