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The dCMP transferase activity of yeast Rev1 is biologically relevant during the bypass of endogenously generated AP sites.

Publication ,  Journal Article
Kim, N; Mudrak, SV; Jinks-Robertson, S
Published in: DNA Repair (Amst)
December 10, 2011

The bypass of AP sites in yeast requires the Rev1 protein in addition to the Pol ζ translesion synthesis DNA polymerase. Although Rev1 was originally characterized biochemically as a dCMP transferase during AP-site bypass, the relevance of this activity in vivo is unclear. The current study uses highly sensitive frameshift- and nonsense-reversion assays to monitor the bypass of AP sites created when uracil is excised from chromosomal DNA. In the frameshift-reversion assay, an unselected base substitution frequently accompanies the selected mutation, allowing the relative incorporation of each of the four dNMPs opposite endogenously created AP sites to be inferred. Results with this assay suggest that dCMP is the most frequent dNMP inserted opposite uracil-derived AP sites and demonstrate that dCMP insertion absolutely requires the catalytic activity of Rev1. In the complementary nonsense-reversion assay, dCMP insertion likewise depended on the dCMP transferase activity of Rev1. Because dAMP insertion opposite uracil-derived AP sites does not revert the nonsense allele and hence could not be detected, it also was possible to detect low levels of dGMP or dTMP insertion upon loss of Rev1 catalytic activity. These results demonstrate that the catalytic activity of Rev1 is biologically relevant and is required specifically for dCMP insertion during the bypass of endogenous AP sites.

Duke Scholars

Published In

DNA Repair (Amst)

DOI

EISSN

1568-7856

Publication Date

December 10, 2011

Volume

10

Issue

12

Start / End Page

1262 / 1271

Location

Netherlands

Related Subject Headings

  • Sequence Deletion
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae
  • Protein Structure, Tertiary
  • Nucleotidyltransferases
  • Molecular Sequence Data
  • Developmental Biology
  • Deoxycytidine Monophosphate
  • DNA-Directed DNA Polymerase
  • DNA, Fungal
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Kim, N., Mudrak, S. V., & Jinks-Robertson, S. (2011). The dCMP transferase activity of yeast Rev1 is biologically relevant during the bypass of endogenously generated AP sites. DNA Repair (Amst), 10(12), 1262–1271. https://doi.org/10.1016/j.dnarep.2011.09.017
Kim, Nayun, Sarah V. Mudrak, and Sue Jinks-Robertson. “The dCMP transferase activity of yeast Rev1 is biologically relevant during the bypass of endogenously generated AP sites.DNA Repair (Amst) 10, no. 12 (December 10, 2011): 1262–71. https://doi.org/10.1016/j.dnarep.2011.09.017.
Kim N, Mudrak SV, Jinks-Robertson S. The dCMP transferase activity of yeast Rev1 is biologically relevant during the bypass of endogenously generated AP sites. DNA Repair (Amst). 2011 Dec 10;10(12):1262–71.
Kim, Nayun, et al. “The dCMP transferase activity of yeast Rev1 is biologically relevant during the bypass of endogenously generated AP sites.DNA Repair (Amst), vol. 10, no. 12, Dec. 2011, pp. 1262–71. Pubmed, doi:10.1016/j.dnarep.2011.09.017.
Kim N, Mudrak SV, Jinks-Robertson S. The dCMP transferase activity of yeast Rev1 is biologically relevant during the bypass of endogenously generated AP sites. DNA Repair (Amst). 2011 Dec 10;10(12):1262–1271.
Journal cover image

Published In

DNA Repair (Amst)

DOI

EISSN

1568-7856

Publication Date

December 10, 2011

Volume

10

Issue

12

Start / End Page

1262 / 1271

Location

Netherlands

Related Subject Headings

  • Sequence Deletion
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae
  • Protein Structure, Tertiary
  • Nucleotidyltransferases
  • Molecular Sequence Data
  • Developmental Biology
  • Deoxycytidine Monophosphate
  • DNA-Directed DNA Polymerase
  • DNA, Fungal