Available online at www.sciencedirect.com ScienceDirect Recent progress in dissecting molecular recognition by DNA polymerases with non-native substrates 1,3 1,2 Kaitlin M Pugliese and Gregory A Weiss DNA polymerases must discriminate the correct Watson–Crick new insights into mechanisms of molecular recognition base pair-forming deoxynucleoside triphosphate (dNTP) essential for the function of DNA polymerase [5 ,8,9]. substrate from three other dNTPs and additional triphosphates found in the cell. The rarity of misincorporations in vivo, then, Non-cognate substrates for DNA polymerization, includ- belies the high tolerance for dNTP analogs observed in ing ribonucleoside triphosphates (rNTPs) and dideoxy- vitro. Advances over the last 10 years in single-molecule nucleoside triphosphates (ddNTPs), represent the ﬂuorescence and electronic detection of dNTP analog molecular recognition challenge faced by DNA polymer- incorporation enable exploration of the mechanism and limits ase. The rNTPs, for example, are present at >10-fold to base discrimination by DNA polymerases. Such studies higher concentrations than dNTPs in the cell , yet are reveal transient motions of DNA polymerase during substrate rejected as substrates by DNA polymerase. Experiments recognition and mutagenesis in the context of erroneous dNTP with such substrates have generated insights into the incorporation that can lead to evolution and genetic disease. enzyme’s
Current Opinion in Chemical Biology – Elsevier
Published: Dec 1, 2017
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