RNase H2-initiated ribonucleotide excision repair

Justin L Sparks; Hyongi Chon; Susana M Cerritelli; Thomas A Kunkel; Erik Johansson; Robert J Crouch; Peter M Burgers

doi:10.1016/j.molcel.2012.06.035

RNase H2-initiated ribonucleotide excision repair

Mol Cell. 2012 Sep 28;47(6):980-6. doi: 10.1016/j.molcel.2012.06.035. Epub 2012 Aug 2.

Authors

Justin L Sparks¹, Hyongi Chon, Susana M Cerritelli, Thomas A Kunkel, Erik Johansson, Robert J Crouch, Peter M Burgers

Affiliation

¹ Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110, USA.

Abstract

Ribonucleotides are incorporated into DNA by the replicative DNA polymerases at frequencies of about 2 per kb, which makes them by far the most abundant form of potential DNA damage in the cell. Their removal is essential for restoring a stable intact chromosome. Here, we present a complete biochemical reconstitution of the ribonucleotide excision repair (RER) pathway with enzymes purified from Saccharomyces cerevisiae. RER is most efficient when the ribonucleotide is incised by RNase H2, and further excised by the flap endonuclease FEN1 with strand displacement synthesis carried out by DNA polymerase δ, the PCNA clamp, its loader RFC, and completed by DNA ligase I. We observed partial redundancy for several of the enzymes in this pathway. Exo1 substitutes for FEN1 and Pol ε for Pol δ with reasonable efficiency. However, RNase H1 fails to substitute for RNase H2 in the incision step of RER.

Publication types

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

MeSH terms

Acetyltransferases / metabolism*
DNA Ligase ATP
DNA Ligases / metabolism
DNA Polymerase II / metabolism
DNA Polymerase III / metabolism
DNA Repair*
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Exodeoxyribonucleases / metabolism
Membrane Proteins / metabolism*
Proliferating Cell Nuclear Antigen / metabolism
Ribonuclease H / metabolism*
Ribonucleotides / metabolism
Saccharomyces cerevisiae / enzymology
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae / metabolism*
Saccharomyces cerevisiae Proteins / metabolism*

Substances

DNA-Binding Proteins
Membrane Proteins
Proliferating Cell Nuclear Antigen
Ribonucleotides
Saccharomyces cerevisiae Proteins
Acetyltransferases
ELO2 protein, S cerevisiae
DNA Polymerase II
DNA Polymerase III
Exodeoxyribonucleases
exodeoxyribonuclease I
ribonuclease HII
Ribonuclease H
ribonuclease HI
DNA Ligases
DNA Ligase ATP

Abstract

Publication types

MeSH terms

Substances

Grants and funding