The consensus sequence for single-base deletions in non-reiterated runs during in vitro DNA-dependent DNA polymerisation is refined using data available in the literature. This leads to the observation that chain termination codons are hotspots for single-base deletions. The evolutionary implications are discussed in two models which differ in whether polymerases evolved while the genetic code emerged or after the genetic code was fixed. A possible answer to the question 'Why are stop codons just what they are?' is suggested.© 1997 Federation of European Biochemical Societies.Key words: Genetic code; Stop codon; Deletion; Polymerase
AssumptionsThe mutational spectra of in vitro polymerisation [1-9] for DNA polymerases belonging to families found in at least two of the three living kingdoms [10] are considered here as relevant with respect to a primordial polymerase; we will not take into account DNA polymerases ß, as they are family X DNA polymerases so far only found among eukaryotes [11], and HIV reverse transcriptases, which emerged very 'late' in evolution [12].As it is believed that RNA preceded DNA in evolution [13], data for RNA replicases would be more relevant but are not available; recent evidence shows, however, that DNA and RNA replicases are very closely related [14][15][16]: a single substitution of a hydroxyl group by a hydrogen atom in the Y639F mutant of T7 RNA polymerase allows a DNA replicase to function as a RNA replicase [17], and a single mutation confers on Moloney murine leukaemia virus reverse transcriptase the ability to replicate RNA [18]; we also note that Escherichia coli DNA polymerase I is an accurate RNA-dependent DNA polymerase [19].
Polymerase errorsPolymerase-induced mutations are mainly substitutions and frameshifts [1][2][3][4][5][6][7]. For the Klenow polymerase domain [1], which has no nuclease domain, as can be assumed for a primordial polymerase, the frameshift error rate is about half the substitution error rate: frameshift mutations therefore represent a significant proportion of the mutations in such systems. Frameshifts result mostly from deletions and additions of one base [1][2][3][4][5]. Crucially, these are highly deleterious by preventing translation in the correct reading frame of the codons »Corresponding author. Fax: (44) (1223) 402 140. E-mail: jlj@mrc-lmb.cam.ac.uk downstream of the mutation. Frameshifts occurring in directly repeated and palindromic sequences [8] will be addressed in the discussion. Here, we will focus on frameshifts in non-reiterated runs, where single-base deletions occur far more frequently than single-base additions [1,2,4,7]. Additions will therefore be neglected in the following. For polymerases with and without nuclease domains, the data indicate no significant differences in the consensus sequence for single-base deletions in non-reiterated runs. It has been defined as YR [1], TTR [9], YTG [6] and TR [8]. Using the current data [1-9] we here refine it as YTRV (V = C, A or G; Table 1). These singlebase deletions are found to...