Differential Activities of DNA Polymerases in Processing Ribonucleotides during DNA Synthesis in Archaea.

摘要: Consistent with the fact that ribonucleotides (rNTPs) are in excess over deoxyribonucleotides (dNTPs) vivo, recent findings indicate replicative DNA polymerases (DNA Pols) able to insert (rNMPs) during synthesis, raising crucial questions about fidelity of replication both Bacteria and Eukarya. Here, we report level rNTPs is 20-fold higher than dNTPs Pyrococcus abyssi cells. Using dNTP rNTP concentrations present recorded rNMP incorporation a template-specific manner vitro family-D Pol (PolD) having highest propensity compared family-B p41/p46 complex. We also showed accumulate at relatively high frequency genome wild-type Thermococcales cells, this significantly increases upon deletion RNase HII, major enzyme responsible for removal RNA from DNA. Because remain genomic DNA, then analyzed effects on polymerization activities by three Pols. Depending identity base sequence context, all Pols bypass rNMP-containing templates variable efficiency nucleotide (mis)incorporation ability. Unexpectedly, found PolD correctly base-paired single ribonucleotide opposite templates. An evolutionary scenario discussed concerning into stability.