DNA-binding proteins as site-specific nucleases.
DNA-binding proteins can be converted into site-specific nucleases by linking them to the chemical nuclease 1,10-phenanthroline-copper. This can be readily accomplished by converting a minor groove-proximal amino acid to a cysteine residue using site-directed mutagenesis and then chemically modifying the sulphydryl group with 5-iodoacetamido-1,10-phenanthroline-copper. These chimeric scission reagents can be used as rare cutters to analyse chromosomal DNA, to test predictions based on high-resolution nuclear magnetic resonance and X-ray crystal structures, and to locate binding sites of proteins within genomes.
Pan
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Molecular Biology Institute, UCLA 90024-1570.
Landgraf
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Sigman
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D002384:Catalysis; D003300:Copper; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D010618:Phenanthrolines
C088903:5-iodoacetamido-1,10-phenanthroline; D004268:DNA-Binding Proteins; D010618:Phenanthrolines; D003300:Copper; D015252:Deoxyribonucleases, Type II Site-Specific; C025205:1,10-phenanthroline