Effect of Variations in the Conditions of DNA Synthesis upon the Accuracy of DNA Replication
Recent refinements in the methods for measurement of accuracy of DNA replication have led to accurate estimates for a number of mispairs during the duplication of natural DNA for bacterial and mammalian DNA polymerases and for E. coli and bacteriophage T4 multienzyme replication complexes (see Refs. 17 and 25 for recent reviews). Using a combination of seven purified replication proteins of phage T4 (the products of genes 32, 41, 43, 44, 45, 61, and 62) to replicate DNA in vitro, it has been shown that the accuracy of this complex is very similar to that seen during replication of ΦX174 DNA in vivo [12, 16, 23–25]. DNA sequences (such as am33) that are replicated with a low accuracy in vivo show high error rates during replication in vitro. Other sequences (such as am16 and am9), which have a low reversion rate in vivo, are replicated with comparably high accuracy in vitro [23–25]. It has also been shown that by the appropriate use of deoxynucleotide pool biases, it is possible to induce reversions of a given amber mutant by specific transition and transversion pathways [12, 23, 24]. We have examined the influence of a variety of perturbations in the conditions of DNA replication in vitro, on the rates of formation of mispairs leading to transition and transversion mutations. These results are summarized here.
KeywordsTransition Pathway Intercalate Agent Base Analog Transversion Mutation Replication Apparatus
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