The DNA polymerase activity of Saccharomyces cerevisiae Rev1 is biologically significant

Genetics. 2011 Jan;187(1):21-35. doi: 10.1534/genetics.110.124172. Epub 2010 Oct 26.

Abstract

A cell's ability to tolerate DNA damage is directly connected to the human development of diseases and cancer. To better understand the processes underlying mutagenesis, we studied the cell's reliance on the potentially error-prone translesion synthesis (TLS), and an error-free, template-switching pathway in Saccharomyces cerevisiae. The primary proteins mediating S. cerevisiae TLS are three DNA polymerases (Pols): Rev1, Pol ζ (Rev3/7), and Pol η (Rad30), all with human homologs. Rev1's noncatalytic role in recruiting other DNA polymerases is known to be important for TLS. However, the biological significance of Rev1's unusual conserved DNA polymerase activity, which inserts dC, is much less well understood. Here, we demonstrate that inactivating Rev1's DNA polymerase function sensitizes cells to both chronic and acute exposure to 4-nitroquinoline-1-oxide (4-NQO) but not to UV or cisplatin. Full Rev1-dependent resistance to 4-NQO, however, also requires the additional Rev1 functions. When error-free tolerance is disrupted through deletion of MMS2, Rev1's catalytic activity is more vital for 4-NQO resistance, possibly explaining why the biological significance of Rev1's catalytic activity has been elusive. In the presence or absence of Mms2-dependent error-free tolerance, the catalytic dead strain of Rev1 exhibits a lower 4-NQO-induced mutation frequency than wild type. Furthermore, Pol ζ, but not Pol η, also contributes to 4-NQO resistance. These results show that Rev1's catalytic activity is important in vivo when the cell has to cope with specific DNA lesions, such as N(2)-dG.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 4-Nitroquinoline-1-oxide / pharmacology
  • Biocatalysis
  • Cisplatin / pharmacology
  • DNA Adducts / genetics
  • DNA Adducts / metabolism
  • DNA Damage
  • DNA Polymerase iota
  • DNA Repair / drug effects
  • DNA Repair / radiation effects
  • DNA, Fungal / biosynthesis
  • DNA, Fungal / genetics
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / metabolism*
  • Humans
  • Mutation / drug effects
  • Mutation / radiation effects
  • Nucleotidyltransferases / chemistry
  • Nucleotidyltransferases / metabolism*
  • Protein Structure, Tertiary
  • Pyruvaldehyde / pharmacology
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / radiation effects
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Ultraviolet Rays

Substances

  • DNA Adducts
  • DNA, Fungal
  • Saccharomyces cerevisiae Proteins
  • 4-Nitroquinoline-1-oxide
  • Pyruvaldehyde
  • DNA polymerase zeta
  • Nucleotidyltransferases
  • REV1 protein, S cerevisiae
  • DNA-Directed DNA Polymerase
  • Cisplatin
  • DNA Polymerase iota