Recognition of HNA and 1,5-anhydrohexitol nucleotides by DNA metabolizing enzymes.
Hexitol nucleic acids (HNA) as well as their 1,5-anhydrohexitol triphosphate building blocks were evaluated for their ability to be recognized by several DNA metabolizing enzymes. It was found that RNA polymerases can recognize the triphosphate of the adenine analogue. However, only the incorporation of a maximum of three consecutive building block analogues was possible under the applied experimental conditions. Terminal transferase was more successful succeeding in the elongation of a DNA primer with a maximum of 15 1,5-anhydrohexitol purine nucleotides. Furthermore, it was observed that the 1,5-anhydroaltritol triphosphate analogue of adenosine was a poor substrate for terminal transferase and that HNA could not act as a primer for this enzyme. Likewise, HNA did not function as a template for restriction enzymes, ligases or methylases.
Vastmans
Karen
K
Laboratory of Medicinal Chemistry, Rega Institute for Medical Research, Katholieke Universiteit Leuven, Minderbroedersstraat 10, B-3000 Louvain, Belgium.
Rozenski
Jef
J
Van Aerschot
Arthur
A
Herdewijn
Piet
P
D000227:Adenine Nucleotides / Q000737:chemistry; D019175:DNA Methylation; D004262:DNA Restriction Enzymes; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry*; D009711:Nucleotides / Q000737:chemistry*; D026723:Polyadenylation; D011685:Purine Nucleotides / Q000737:chemistry; D013402:Sugar Alcohols / Q000737:chemistry*; D014158:Transcription, Genetic
D000227:Adenine Nucleotides; D009711:Nucleotides; D011685:Purine Nucleotides; D013402:Sugar Alcohols; D012321:DNA-Directed RNA Polymerases; D004262:DNA Restriction Enzymes