Ribonucleotides as nucleotide excision repair substrates

Research output: Contribution to journalArticle

Abstract

The incorporation of ribonucleotides in DNA has attracted considerable notice in recent years, since the pool of ribonucleotides can exceed that of the deoxyribonucleotides by at least 10-20-fold, and single ribonucleotide incorporation by DNA polymerases appears to be a common event. Moreover ribonucleotides are potentially mutagenic and lead to genome instability. As a consequence, errantly incorporated ribonucleotides are rapidly repaired in a process dependent upon RNase H enzymes. On the other hand, global genomic nucleotide excision repair (NER) in prokaryotes and eukaryotes removes damage caused by covalent modifications that typically distort and destabilize DNA through the production of lesions derived from bulky chemical carcinogens, such as polycyclic aromatic hydrocarbon metabolites, or via crosslinking. However, a recent study challenges this lesion-recognition paradigm. The work of Vaisman et al. (2013) [34] reveals that even a single ribonucleotide embedded in a deoxyribonucleotide duplex is recognized by the bacterial NER machinery in vitro. In their report, the authors show that spontaneous mutagenesis promoted by a steric-gate pol V mutant increases in uvrA, uvrB, or uvrC strains lacking rnhB (encoding RNase HII) and to a greater extent in an NER-deficient strain lacking both RNase HI and RNase HII. Using purified UvrA, UvrB, and UvrC proteins in in vitro assays they show that despite causing little distortion, a single ribonucleotide embedded in a DNA duplex is recognized and doubly-incised by the NER complex. We present the hypothesis to explain the recognition and/or verification of this small lesion, that the critical 2'-OH of the ribonucleotide - with its unique electrostatic and hydrogen bonding properties - may act as a signal through interactions with amino acid residues of the prokaryotic NER complex that are not possible with DNA. Such a mechanism might also be relevant if it were demonstrated that the eukaryotic NER machinery likewise incises an embedded ribonucleotide in DNA.

Original languageEnglish (US)
Pages (from-to)55-60
Number of pages6
JournalDNA Repair
Volume13
Issue number1
DOIs
StatePublished - Jan 2014

Fingerprint

Ribonucleotides
DNA Repair
Repair
Nucleotides
Substrates
Deoxyribonucleotides
DNA
Machinery
Ribonuclease H
Mutagenesis
Genomic Instability
Polycyclic Aromatic Hydrocarbons
DNA-Directed DNA Polymerase
Hydrogen Bonding
Metabolites
Eukaryota
Static Electricity
Carcinogens
Crosslinking
Electrostatics

Keywords

  • Electrostatic and hydrogen bonding properties
  • Lesion recognition and verification
  • Prokaryotic nucleotide excision repair
  • Ribonucleotides

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Ribonucleotides as nucleotide excision repair substrates. / Cai, Yuqin; Geacintov, Nicholas; Broyde, Suse.

In: DNA Repair, Vol. 13, No. 1, 01.2014, p. 55-60.

Research output: Contribution to journalArticle

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