Altered nucleotide misinsertion fidelity associated with polι-dependent replication at the end of a DNA template

Ekaterina G. Frank, Agnès Tissier, John P. McDonald, Vesna Rapić-Otrin, Xianmin Zeng, Patricia J. Gearhart, Roger Woodgate

Research output: Contribution to journalArticlepeer-review

Abstract

A hallmark of human DNA polymerase ι (polι) is the asymmetric fidelity of replication at template A and T when the enzyme extends primers annealed to a single-stranded template. Here, we report on the efficiency and accuracy of polι-dependent replication at a nick, a gap, the very end of a template and from a mispaired primer. Polι cannot initiate synthesis on a nicked DNA substrate, but fills short gaps efficiently. Surprisingly, polι's ability to blunt-end a 1 bp recessed terminus is dependent upon the template nucleotide encountered and is highly erroneous. At template G, both C and T are inserted with roughly equal efficiency, whilst at template C, C and A are misinserted 8- and 3-fold more often than the correct base, G. Using substrates containing mispaired primer termini, we show that polι can extend all 12 mispairs, but with differing efficiencies. Polι can also extend a tandem mispair, especially when it is located within a short gap. The enzymatic properties of polι appear consistent with that of a somatic hypermutase and suggest that polι may be one of the low-fidelity DNA polymerases hypothesized to participate in the hypermutation of immunoglobulin variable genes in vivo.

Original languageEnglish (US)
Pages (from-to)2914-2922
Number of pages9
JournalEMBO Journal
Volume20
Issue number11
DOIs
StatePublished - Jun 1 2001

Keywords

  • DNA polymerase ζ
  • DNA polymerase η
  • Rad30
  • Rad30B
  • Somatic mutation

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Fingerprint Dive into the research topics of 'Altered nucleotide misinsertion fidelity associated with polι-dependent replication at the end of a DNA template'. Together they form a unique fingerprint.

Cite this