NDLI: Mutational clusters generated by non-processive polymerases: a case study using DNA polymerase ß in vitro

Content Provider	PubMed Central
Author	Libertad, GarcÃa-villada Drake, John W.
Abstract	Available DNA mutational spectra reveal that the number of mutants with multiple mutations (“multiples”) is usually greater than expected from a random distribution of mutations among mutants. These overloads imply the occurrence of non-random clusters of mutations, probably generated during episodes of low-fidelity DNA synthesis. Excess multiples have been reported not only for viruses, bacteria, and eukaryotic cells but also for the DNA polymerases of phages T4 and RB69 in vitro. In the simplest case of a purified polymerase, non-random clusters may be generated by a subfraction of phenotypic variants able to introduce more errors per cycle of DNA synthesis than the normal enzyme. According to this hypothesis, excess multiples are not expected with non-processive polymerases even if they harbor rare mutator variants. DNA polymerase ß (Pol ß) is a mammalian DNA-repair polymerase with very low processivity. Although several Pol ß mutational spectra have been described, there is conflicting evidence on whether or not excess multiples occur, with spectra based on the HSV-tk system tending to show excess multiples. Excess multiples generated by Pol ß or any of its mutants might imply that the excesses of multiples observed in numerous other systems, especially those with processive polymerases, could be artifactual. Here, the distributions of mutations generated by native and recombinant rat Pol ß and by the Pol ßY265C mutator were analyzed in the M13mp2 lacZα system. Our results present no evidence for a significant excess of multiples over the expected numbers with any of the Pol ß enzymes tested in this system. The reported excess of Pol ß-generated multiples in the HSV-tk system may reflect a reduced efficiency of detection of base substitutions that cause weak phenotypes, which in turn may artifactually increase the frequency of multiples.
Related Links	http://dx.doi.org/10.1016/j.dnarep.2010.05.002
Ending Page	878
Page Count	8
Starting Page	871
File Format	PDF
ISSN	15687864
e-ISSN	15687856
Journal	DNA repair
Issue Number	8
Volume Number	9
Language	English
Publisher Date	2010-08-01
Access Restriction	Open
Subject Keyword	Cell Biology Biochemistry Molecular Biology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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