Ribonuclease H2, subunit B is a protein that in humans is encoded by the RNASEH2B gene. RNase H2 is composed of a single catalytic subunit (A) and two non-catalytic subunits (B and C), and degrades the RNA of RNA:DNA hybrids. The non-catalytic B subunit of RNase H2 is thought to play a role in DNA replication.
|Recessive lethal study||Abnormal|
|Glucose tolerance test||Normal|
|Auditory brainstem response||Normal|
|All tests and analysis from|
Model organisms have been used in the study of RNASEH2B function. A conditional knockout mouse line, called Rnaseh2btm1a(EUCOMM)Wtsi was generated as part of the International Knockout Mouse Consortium program — a high-throughput mutagenesis project to generate and distribute animal models of disease to interested scientists.
Male and female animals underwent a standardized phenotypic screen to determine the effects of deletion. Twenty four tests were carried out on mutant mice and three significant abnormalities were observed. No homozygous mutant embryos were identified during gestation, and therefore none survived until weaning. The remaining tests were carried out on heterozygous mutant adult mice and an increased susceptibility to bacterial infection was observed in female animals.
In a study of Reijns et al. 2012, it was made targeted mutagenesis of the mouse Rnaseh2b gene to gain insight into the in vivo role of the mammalian RNase H2 enzyme, due to the fact that the ablation of Rnaseh2b in mice leads to early embryonic lethality. Their hypothesis was that the growth arrest was a consecuence of a p53-dependent DNA damage response associated with the accumulation of single RN in genomic DNA.
They discuss the following facts:
1. Ribonucleotide accumulate in RNaseH2 null cells as a consequence of incorporation by DNA polymerases: The authors show that the ribonucleotide incorporation also occurs in metazoans, these lesions are harmful to mammalian cells, and their removal is required for mouse embryonic development. They also characterize alkali-sensitive sites: Lesions are single or diRN covalently incorporated into genomic DNA, at a frequency of approx. 1.000.000 sites per cell, making it the most common endogenous base lesion in the mammalian genome. These lesions are best explained by misincorporation by the major replicative polymerases.
2. RNase H2 is a genome surveillance enzyme required for ribonucleotide removal: They found that ribonucleotide accumulation in genomic DNA of RNaseH2null mice implicates the RNaseH2 complex in the maintenance of genome integrity. These ribonucleotide changes are likely to be harmful, as their ribose 2’-hydroxyl group increases susceptibility of the adjacent phosphodiester bond to hydrolysis. Actually, they report that the ribonucleotides are being incorporated 1 every ~7.600 nt in null cells = 1.300.000 lesions per cell. This have the same order of magnitude predicted from in vitro incorporation rates by eukaryotic replicative polymerases.
3. Misincorporated ribonucleotide induce DNA damage: It´s not that the ribonucleotides do not prevent replication; actually, polDNA can tolerate templates with ribonucleotides, having normal early embryogenesis. The problem appears with excessive numbers of ribonucleotides. DNA damage response signaling activated maybe by incorporation of ribonucleotides in difficult to replicate regions or near other detrimental lesions. They also found chromosomal rearrangements: DNA breaks may origin by replication fork collapse or hydrolysis of RN on opposing DNA strands. Also, the marked activation of DNA damage signaling in embryo may produce a p53-mediated inhibition of proliferation that might contribute to the lethality of null embryos.
4. Ribonucleotide incorporation in health and disease: Past studies reported only two contexts where there’s stable incorporation of ribonucleotides: 1) diRibonucleotides in S. pombe may be a signal to initiate homologous recombination. 2) Ribonucleotides in mtDNA (Mouse and HeLa cells). Low levels of ribonucleotides incorporation in the nuclear genome may be tolerated. Actually, aberrant nucleic acid substrates, generated by repair pathways non-RNaseH2 dependent (due to reduced RNaseH2 activity in Aicardi-Goutières Syndrome) are thought to drive innate immune response. Alternatively, ribonucleotides might induce DNA-damage response signaling that by itself may stimulates interferon production.
In summary, this study highlights the fact that the ribonucleotides can be highly deleterious to the mammalian cell, causing genome instability, and that the RNaseH2 is a critical enzyme for ensuring integrity of genomic DNA. It also calls for attention and interest about the pathway(s) that remove ribonucleotides from genomic DNA, site and nature of ribonucleotides -induced DNA damage, and distribution of ribonucleotides in the genome. Knowing this, it may gain understanding about the pathological and physiological roles of RN in genomic DNA, of significance to both nucleic acid-driven autoimmunity and carcinogenesis.
- "ribonuclease H2, subunit B". Retrieved 2011-12-04.
- Crow, Y. J.; Leitch, A.; Hayward, B. E.; Garner, A.; Parmar, R.; Griffith, E.; Ali, M.; Semple, C.; Aicardi, J.; Babul-Hirji, R.; Baumann, C.; Baxter, P.; Bertini, E.; Chandler, K. E.; Chitayat, D.; Cau, D.; Déry, C.; Fazzi, E.; Goizet, C.; King, M. D.; Klepper, J.; Lacombe, D.; Lanzi, G.; Lyall, H.; Martínez-Frías, M. A. L.; Mathieu, M. L.; McKeown, C.; Monier, A.; Oade, Y.; Quarrell, O. W. (2006). "Mutations in genes encoding ribonuclease H2 subunits cause Aicardi-Goutières syndrome and mimic congenital viral brain infection". Nature Genetics 38 (8): 910–916. doi:10.1038/ng1842. PMID 16845400.
- "Salmonella infection data for Rnaseh2b". Wellcome Trust Sanger Institute.
- "Citrobacter infection data for Rnaseh2b". Wellcome Trust Sanger Institute.
- Gerdin AK (2010). "The Sanger Mouse Genetics Programme: High throughput characterisation of knockout mice". Acta Ophthalmologica 88: 925–7. doi:10.1111/j.1755-3768.2010.4142.x.
- Mouse Resources Portal, Wellcome Trust Sanger Institute.
- "International Knockout Mouse Consortium".
- "Mouse Genome Informatics".
- Skarnes, W. C.; Rosen, B.; West, A. P.; Koutsourakis, M.; Bushell, W.; Iyer, V.; Mujica, A. O.; Thomas, M.; Harrow, J.; Cox, T.; Jackson, D.; Severin, J.; Biggs, P.; Fu, J.; Nefedov, M.; De Jong, P. J.; Stewart, A. F.; Bradley, A. (2011). "A conditional knockout resource for the genome-wide study of mouse gene function". Nature 474 (7351): 337–342. doi:10.1038/nature10163. PMC 3572410. PMID 21677750.
- Dolgin E (2011). "Mouse library set to be knockout". Nature 474 (7351): 262–3. doi:10.1038/474262a. PMID 21677718.
- Collins FS, Rossant J, Wurst W (2007). "A Mouse for All Reasons". Cell 128 (1): 9–13. doi:10.1016/j.cell.2006.12.018. PMID 17218247.
- van der Weyden L, White JK, Adams DJ, Logan DW (2011). "The mouse genetics toolkit: revealing function and mechanism.". Genome Biol 12 (6): 224. doi:10.1186/gb-2011-12-6-224. PMC 3218837. PMID 21722353.
- Reijns, Martin; Björn Rabe, Rachel E. Rigby, Pleasantine Mill, Katy R. Astell, Laura A. Lettice, Shelagh Boyle, Andrea Leitch, Margaret Keighren, Fiona Kilanowski, Paul S. Devenney, David Sexton, Graeme Grimes, Ian J. Holt, Robert E. Hill, Martin S. Taylor, Kirstie A. Lawson, Julia R. Dorin, Andrew P. Jackson (25 May 2012). "Enzymatic Removal of Ribonucleotides from DNA Is Essential for Mammalian Genome Integrity and Development". Cell 149 (5): 1008. doi:10.1016/j.cell.2012.04.011. Retrieved 18 January 2013.
- Chon, H.; Vassilev, A.; Depamphilis, M. L.; Zhao, Y.; Zhang, J.; Burgers, P. M.; Crouch, R. J.; Cerritelli, S. M. (2008). "Contributions of the two accessory subunits, RNASEH2B and RNASEH2C, to the activity and properties of the human RNase H2 complex". Nucleic Acids Research 37 (1): 96–110. doi:10.1093/nar/gkn913. PMC 2615623. PMID 19015152.
- Crow, Y. J.; Livingston, J. H. (2008). "Aicardi-Goutières syndrome: An important Mendelian mimic of congenital infection". Developmental Medicine & Child Neurology 50 (6): 410–416. doi:10.1111/j.1469-8749.2008.02062.x. PMID 18422679.
- Ali, M.; Highet, L. J.; Lacombe, D.; Goizet, C.; King, M. D.; Tacke, U.; Van Der Knaap, M. S.; Lagae, L.; Rittey, C.; Brunner, H. G.; Van Bokhoven, H.; Hamel, B.; Oade, Y. A.; Sanchis, A.; Desguerre, I.; Cau, D.; Mathieu, N.; Moutard, M. L.; Lebon, P.; Kumar, D.; Jackson, A. P.; Crow, Y. J. (2005). "A second locus for Aicardi‐Goutières syndrome at chromosome 13q14–21". Journal of Medical Genetics 43 (5): 444–450. doi:10.1136/jmg.2005.031880. PMC 2649012. PMID 15908569.