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Stimulation of human flap endonuclease 1 by human immunodeficiency virus type 1 integrase: Possible role for flap endonuclease 1 in 5′-end processing of human immunodeficiency virus type 1 integration intermediates

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Human immunodeficiency virus type 1 (HIV-1) DNA integration intermediates consist of viral and host DNA segments separated by a 5-nucleotide gap adjacent to a 5′-AC unpaired dinucleotide. These short-flap(pre-repair) integration intermediates are structurally similar to DNA loci undergoing long-patch base excision repair in mammalian cells. The cellular proteins flap endonuclease 1 (FEN-1), proliferating cell nuclear antigen, replication factor C, DNA ligase I and DNA polymerase delta are required for the repair of this type of DNA lesion. The role of FEN-1 in the base excision repair pathway is to cleave 5′-unpaired flaps in forked structures so that DNA ligase can seal the single-stranded breaks that remain following gap repair. The rate of excision by FEN-1 of 5′-flaps from short- and long-flap oligonucleotide substrates that mimicpre- andpost-repair HIV-1 integration intermediates, respectively, and the effect of HIV-1 integrase on these reactions were examined in the present study. Cleavage of 5′-flaps by FEN-1 inpre-repair HIV-1 integration intermediates was relatively inefficient and was further decreased 3-fold by HIV-1 integrase. The rate of removal of 5′-flaps by FEN-1 frompost-repair HIV-1 integration intermediates containing relatively long (7-nucleotide) unpaired 5′-tails and short (1-nucleotide) gaps was increased 3-fold relative to that seen withpre-repair substrates and was further stimulated 5- to 10-fold by HIV-1 integrase. Overall,post-repair structures were cleaved 18 times more effectively in the presence of HIV-1 integrase thanpre-repair structures. The site of cleavage was 1 or 2 nucleotides 3′ of the branch point and was unaffected by HIV-1 integrase. Integrase alone had no detectable activity in removing 5′-flaps from eitherpre- orpost-repair substrates.

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Faust, E.A., Triller, H. Stimulation of human flap endonuclease 1 by human immunodeficiency virus type 1 integrase: Possible role for flap endonuclease 1 in 5′-end processing of human immunodeficiency virus type 1 integration intermediates. J Biomed Sci 9, 273–287 (2002).

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Key Words

  • HIV-1
  • Integration
  • DNA repair
  • FEN-1