Human DNA polymerase κ forms nonproductive complexes with matched primer termini but not with mismatched primer termini

Karissa D. Carlson, Robert E. Johnson, Louise Prakash, Satya Prakash, M. Todd Washington

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Human DNA polymerase kappa (pol κ) is a member of the Y family of DNA polymerases that function in translesion synthesis. It synthesizes DNA with moderate fidelity and does not efficiently incorporate nucleotides opposite DNA lesions. Pol κ has the unusual ability to efficiently extend from mismatched primer termini, and it extends readily from nucleotides inserted by other DNA polymerases opposite a variety of DNA lesions. All of this has suggested that pol κ functions during the extension step of translesion synthesis. Here, we have carried out pre-steady-state kinetic studies of pol κ using DNA with matched and mismatched primer termini. Interestingly, we find that mismatches present only a modest kinetic barrier to nucleotide incorporation by pol κ. Moreover, and quite surprisingly, active-site titrations revealed that the concentration of active pol κ is very low with matched DNA, and from DNA trapping experiments we determined that this was due to the formation of nonproductive protein-DNA complexes. In marked contrast, we found that the concentration of active pol κ was six-fold greater with mismatched DNA than with matched DNA. Thus, pol κ forms nonproductive complexes with matched but not with mismatched DNA. From these observations, we conclude that pol κ has evolved to specifically function on DNA substrates with aberrant primer-terminal base pairs, such as the ones it would encounter during the extension step of translesion synthesis.

Original languageEnglish (US)
Pages (from-to)15776-15781
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number43
DOIs
StatePublished - Oct 24 2006

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DNA-Directed DNA Polymerase
DNA
Nucleotides
Aptitude
Base Pairing
Catalytic Domain

Keywords

  • DNA damage
  • DNA repair
  • DNA replication
  • Kinetics
  • Mutagenesis

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Human DNA polymerase κ forms nonproductive complexes with matched primer termini but not with mismatched primer termini. / Carlson, Karissa D.; Johnson, Robert E.; Prakash, Louise; Prakash, Satya; Washington, M. Todd.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 103, No. 43, 24.10.2006, p. 15776-15781.

Research output: Contribution to journalArticle

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