5-amino-2'-deoxyuridine, a novel thymidine analogue for high-resolution footprinting of protein-DNA complexes.
[formula: see text] 5-Amino-2'-deoxyuridine 5'-triphosphate, an analogue of deoxythymidine triphosphate, was synthesized and found to be a substrate of Taq DNA polymerase. The DNA-borne analogue underwent selective chemical reaction with permanganate. The use of 5-amino-dU as an interference probe was validated using the Ada protein/ada promoter complex. The performance of 5-amino-dU in interference footprints is similar to that of the previously described analogue 5-hydroxy-dU, but the former is incorporated more readily into DNA during enzymatic polymerization.
Storek
Michael J
MJ
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Suciu
Alexandru
A
Verdine
Gregory L
GL
D001426:Bacterial Proteins / Q000737:chemistry; D001665:Binding Sites; D004247:DNA / Q000378:metabolism; D018983:DNA Footprinting / Q000379:methods*; D004268:DNA-Binding Proteins / Q000737:chemistry; D003857:Deoxyuridine / Q000031:analogs & derivatives*; D029968:Escherichia coli Proteins; D015335:Molecular Probes; D019853:O(6)-Methylguanine-DNA Methyltransferase; D011485:Protein Binding; D012680:Sensitivity and Specificity; D014157:Transcription Factors
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D015335:Molecular Probes; D014157:Transcription Factors; C410463:5-amino-2'-deoxyuridine; D004247:DNA; C045675:Ada protein, E coli; D019853:O(6)-Methylguanine-DNA Methyltransferase; D003857:Deoxyuridine