Enzymatic Removal of Ribonucleotides from DNA Is Essential for Mammalian Genome Integrity and Development

Reijns, Martin A.M.; Rabe, Björn; Rigby, Rachel E.; Mill, Pleasantine; Astell, Katy R.; Lettice, Laura A.; Boyle, Shelagh; Leitch, Andrea; Keighren, Margaret; Kilanowski, Fiona; Devenney, Paul S.; Sexton, David; Grimes, Graeme R.; Holt, Ian; Hill, Robert E.; Taylor, Martin S.; Lawson, Kirstie A.; Dorin, Julia R.; Jackson, Andrew P.

doi:10.1016/j.cell.2012.04.011

Cited by 397 publications

(599 citation statements)

References 89 publications

Supporting

Mentioning

566

Contrasting

Unclassified

Order By: Relevance

“…

Validation of Rnaseh2b −/− MEF lines. RER activity (DRD:DNA) is undetectable in a Rnaseh2b −/− line consistent with complete inactivation of the Rnaseh2b gene (Reijns et al , 2012). Mean of three independent experiments; error bars, SEM, **** P < 0.0001, two‐tailed t ‐test.

…”

Section: Resultsmentioning

confidence: 84%

“…To address this, we examined cells from a second independent mouse line carrying a null allele of Rnaseh2b ( Rnaseh2b tm1d ) , derived from the EUCOMM knockout‐first Rnaseh2b tm1a allele, from which exon 5 of Rnaseh2b had been excised by Cre recombinase. Given the early embryonic lethality of Rnaseh2b −/− mice (Reijns et al , 2012), mouse embryonic fibroblasts (MEFs) were generated on a p53 −/− background. Hereafter, we will refer to the resulting Rnaseh2b −/− p53 −/− and Rnaseh2b +/+ p53 −/− cells simply as Rnaseh2b −/− and Rnaseh2b +/+ , respectively.…”

Section: Resultsmentioning

confidence: 99%

“…Abrogated RNase H2 function could give rise to cytosolic accumulation of RNA:DNA heteroduplexes as a consequence of reduced RNA:DNA degradation of structures such as R‐loops or active retroelements/endogenous retroviruses (Chon et al , 2013; Rigby et al , 2014; Moelling & Broecker, 2015). Alternatively, cytoplasmic DNA with embedded ribonucleotides may accumulate as a consequence of impaired RER that resulted in genome instability (Reijns et al , 2012; Hiller et al , 2012). To address these possibilities, we performed complementation experiments in RNase H2 null cells, to establish which enzymatic activity was associated with the pro‐inflammatory response.…”

Section: Resultsmentioning

confidence: 99%

“…All genotyping PCRs were performed using a multiplex three‐primer strategy and Taq ReddyMix PCR Master Mix (Thermo Scientific), as previously described for Rnaseh2b E202X/E202X (Reijns et al , 2012) and p53 −/− (Jacks et al, 1994). For Rnaseh2b tm1d/+ , Sting/Mpys +/ − as well as all other primers and product sizes, see Appendix Table S2.…”

Section: Methodsmentioning

confidence: 99%

“…Unlike RNase H1, RNase H2 also cleaves and initiates the removal of single ribonucleotides embedded in DNA (Eder et al , 1993; Rydberg & Game, 2002), a process known as ribonucleotide excision repair (RER) (Sparks et al , 2012). RNase H2 is essential for mammalian genome stability, with complete loss of RNase H2 resulting in embryonic lethality (Reijns et al , 2012; Hiller et al , 2012). …”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Ribonuclease H2 mutations induce a cGAS/STING‐dependent innate immune response

et al. 2016

Self Cite

View full text Add to dashboard Cite

Aicardi–Goutières syndrome (AGS) provides a monogenic model of nucleic acid‐mediated inflammation relevant to the pathogenesis of systemic autoimmunity. Mutations that impair ribonuclease (RNase) H2 enzyme function are the most frequent cause of this autoinflammatory disorder of childhood and are also associated with systemic lupus erythematosus. Reduced processing of either RNA:DNA hybrid or genome‐embedded ribonucleotide substrates is thought to lead to activation of a yet undefined nucleic acid‐sensing pathway. Here, we establish Rnaseh2b A174T/A174T knock‐in mice as a subclinical model of disease, identifying significant interferon‐stimulated gene (ISG) transcript upregulation that recapitulates the ISG signature seen in AGS patients. The inflammatory response is dependent on the nucleic acid sensor cyclic GMP‐AMP synthase (cGAS) and its adaptor STING and is associated with reduced cellular ribonucleotide excision repair activity and increased DNA damage. This suggests that cGAS/STING is a key nucleic acid‐sensing pathway relevant to AGS, providing additional insight into disease pathogenesis relevant to the development of therapeutics for this childhood‐onset interferonopathy and adult systemic autoimmune disorders.

show abstract

“…

…”

Section: Resultsmentioning

confidence: 84%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Ribonuclease H2 mutations induce a cGAS/STING‐dependent innate immune response

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

Ribonucleotides in mitochondrial DNA

Wanrooij

Chabes

2019

FEBS Letters

View full text Add to dashboard Cite

The incorporation of ribonucleotides (rNMPs) into DNA during genome replication has gained substantial attention in recent years and has been shown to be a significant source of genomic instability. Studies in yeast and mammals have shown that the two genomes, the nuclear DNA (nDNA) and the mitochondrial DNA (mtDNA), differ with regard to their rNMP content. This is largely due to differences in rNMP repair – whereas rNMPs are efficiently removed from the nuclear genome, mitochondria lack robust mechanisms for removal of single rNMPs incorporated during DNA replication. In this minireview, we describe the processes that determine the frequency of rNMPs in the mitochondrial genome and summarise recent findings regarding the effect of incorporated rNMPs on mtDNA stability and function.

show abstract

Eukaryotic DNA Polymerases

Cotterill

Kearsey

2014

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Deoxyribonucleic acid (DNA) is replicated and repaired by a family of enzymes called DNA polymerases. Eukaryotic cells have a diversity of these enzymes that, while sharing a common biochemical activity, are specialised for particular roles. Three polymerases are required for the replication of the nuclear genome, with Pol α involved in priming and initial synthesis and Pols δ and ϵ involved in bulk DNA replication. These polymerases are dependent on a large number of other proteins which unwind the DNA and perform other functions essential for efficient DNA synthesis. Polymerases are also involved in DNA repair and many repair‐specific enzymes have been identified. Some repair polymerases can refill a gap generated by removal of damaged DNA, or copy a damaged template, allowing DNA synthesis to proceed across a damaged template. Repair polymerases can also have tissue‐specific functions in lymphoid cells, where they contribute to somatic hypermutation of immunoglobulin genes. Key Concepts: Catalytic function of DNA polymerases. Concepts of ‘proofreading’ and ‘processivity’. Roles of replicative DNA polymerases needed for chromosome replication and organisation at the replication fork. Function of accessory proteins needed for polymerase function in chromosome replication. Different modes of action of specialised polymerases involved in DNA repair. Catalytic mechanism of polymerases. Assays used to detect polymerase function in vitro . Relevance of DNA polymerases to human disease.

show abstract

Enzymatic Removal of Ribonucleotides from DNA Is Essential for Mammalian Genome Integrity and Development

Cited by 397 publications

References 89 publications

Ribonuclease H2 mutations induce a cGAS/STING‐dependent innate immune response

Ribonuclease H2 mutations induce a cGAS/STING‐dependent innate immune response

Ribonucleotides in mitochondrial DNA

Eukaryotic DNA Polymerases

Contact Info

Product

Resources

About