DNA polymerase kappa microsatellite synthesis: Two distinct mechanisms of slippage-mediated errors

Beverly A. Baptiste, Kristin Eckert

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

Microsatellite tandem repeats are frequent sites of strand slippage mutagenesis in the human genome. Microsatellite mutations often occur as insertion/deletion of a repeat motif (unit-based indels), and increase in frequency with increasing repeat length after a threshold is reached. We recently demonstrated that DNA polymerase κ (Pol κ) produces fewer unit-based indel errors within dinucleotide microsatellites than does polymerase δ. Here, we examined human Pol κ's error profile within microsatellite alleles of varying sequence composition and length, using an in vitro HSV-tk gap-filling assay. We observed that Pol κ displays relatively accurate synthesis for unit-based indels, using di- and tetranucleotide repeat templates longer than the threshold length. We observed an abrupt increase in the unit-based indel frequency when the total microsatellite length exceeds 28 nucleotides, suggesting that extended Pol κ protein-DNA interactions enhance fidelity of the enzyme when synthesizing these microsatellite alleles. In contrast, Pol κ is error-prone within the HSV-tk coding sequence, producing frequent single-base errors in a manner that is highly biased with regard to sequence context. Single-nucleotide errors are also created by Pol κ within di- and tetranucleotide repeats, independently of the microsatellite allele length and at a frequency per nucleotide similar to the frequency of single base errors within the coding sequence. These single-base errors represent the mutational signature of Pol κ, and we propose them a mechanism independent of homology-stabilized slippage. Pol κ's dual fidelity nature provides a unique research tool to explore the distinct mechanisms of slippage-mediated mutagenesis.

Original languageEnglish (US)
Pages (from-to)787-796
Number of pages10
JournalEnvironmental and Molecular Mutagenesis
Volume53
Issue number9
DOIs
StatePublished - Dec 1 2012

All Science Journal Classification (ASJC) codes

  • Epidemiology
  • Genetics(clinical)
  • Health, Toxicology and Mutagenesis

Fingerprint Dive into the research topics of 'DNA polymerase kappa microsatellite synthesis: Two distinct mechanisms of slippage-mediated errors'. Together they form a unique fingerprint.

  • Cite this