Yeast Tdp1 and Rad1-Rad10 function as redundant pathways for repairing Top1 replicative damage

Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13669-74. doi: 10.1073/pnas.202242599. Epub 2002 Oct 4.

Abstract

When a replication fork collides with a DNA topoisomerase I (Top1) cleavage complex, the covalently bound enzyme must be removed from the DNA 3' end before recombination-dependent replication restart. Here we report that the tyrosyl-DNA phosphodiesterase Tdp1 and the structure-specific endonuclease Rad1-Rad10 function as primary alternative pathways of Top1 repair in Saccharomyces cerevisiae. Thus, tdp1 rad1 cells (including the catalytic point mutant rad1-D869A) not only are highly sensitive to the Top1 poison camptothecin but also exhibit a TOP1-dependent growth delay. Extensive genetic analysis revealed that both Tdp1 and Rad1-Rad10 repair proceed through recombination that equally depends on RAD52, RAD51, and RAD50. The Rad1-Rad10 pathway further particularly depends on RAD59 and SRS2 but is independent of other nucleotide excision repair genes. Although this pattern is consistent with Rad1-Rad10 removing Top1 in a manner similar to its removal of nonhomologous tails during gene conversion, these differ in that Top1 removal does not require Msh2-Msh3. Finally, we show that yeast lacking the Rad1-Rad10-related proteins Mus81-Mms4 display a unique pattern of camptothecin sensitivity and suggest a concerted model for the action of these endonucleases.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Camptothecin / pharmacology
  • DNA Damage
  • DNA Repair Enzymes
  • DNA Repair* / drug effects
  • DNA Repair* / genetics
  • DNA Replication
  • DNA Topoisomerases, Type I / metabolism*
  • DNA, Fungal / metabolism
  • DNA-Binding Proteins / metabolism
  • Endonucleases / metabolism*
  • Enzyme Inhibitors / pharmacology
  • Flap Endonucleases
  • Fungal Proteins / metabolism*
  • Genes, Fungal
  • Models, Biological
  • Mutation
  • Oligonucleotide Array Sequence Analysis
  • Phosphoric Diester Hydrolases / metabolism*
  • Recombination, Genetic
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins*
  • Single-Strand Specific DNA and RNA Endonucleases
  • Topoisomerase I Inhibitors
  • Trans-Activators / metabolism

Substances

  • DNA, Fungal
  • DNA-Binding Proteins
  • Enzyme Inhibitors
  • Fungal Proteins
  • Saccharomyces cerevisiae Proteins
  • Topoisomerase I Inhibitors
  • Trans-Activators
  • Endonucleases
  • Flap Endonucleases
  • MUS81 protein, S cerevisiae
  • RAD1 protein, S cerevisiae
  • MMS4 protein, S cerevisiae
  • RAD10 protein, S cerevisiae
  • Single-Strand Specific DNA and RNA Endonucleases
  • Phosphoric Diester Hydrolases
  • tyrosyl-DNA phosphodiesterase
  • DNA Topoisomerases, Type I
  • DNA Repair Enzymes
  • Camptothecin