RAD18

RAD18 E3 ubiquitin protein ligase
Available structures PDB Ortholog search: PDBe, RCSB List of PDB id codes 2MRE​, 2MRF​, 2Y43​, 2YBF​
Identifiers
Symbols	RAD18 ; RNF73
External IDs	OMIM: 605256 MGI: 1890476 HomoloGene: 48572 GeneCards: RAD18 Gene
Gene ontology Molecular function • Y-form DNA binding • damaged DNA binding • protein binding • zinc ion binding • ligase activity • polyubiquitin binding • ubiquitin protein ligase binding • single-stranded DNA-dependent ATPase activity • ubiquitin protein ligase activity Cellular component • chromatin • XY body • nucleus • nucleoplasm • replication fork • Rad6-Rad18 complex Biological process • DNA repair • postreplication repair • protein monoubiquitination • spermatogenesis • response to UV • translesion synthesis • DNA damage response, detection of DNA damage • negative regulation of DNA recombination Sources: Amigo / QuickGO
Orthologs
Species	Human	Mouse
Entrez	56852	58186
Ensembl	ENSG00000070950	ENSMUSG00000030254
UniProt	Q9NS91	Q9QXK2
RefSeq (mRNA)	NM_020165	NM_001167730
RefSeq (protein)	NP_064550	NP_001161202
Location (UCSC)	Chr 3: 8.78 – 8.96 Mb	Chr 6: 112.62 – 112.7 Mb
PubMed search

E3 ubiquitin-protein ligase RAD18 is an enzyme that in humans is encoded by the RAD18 gene.^[1][2][3]

The protein encoded by this gene is highly similar to S. cerevisiae DNA damage repair protein Rad18. Yeast Rad18 functions through its interaction with Rad6, which is a ubiquitin-conjugating enzyme required for post-replication repair of damaged DNA. Similar to its yeast counterpart, this protein is able to interact with the human homolog of yeast Rad6 protein through a conserved ring finger motif. Mutation of this motif results in defective replication of UV-damaged DNA and hypersensitivity to multiple mutagens.^[3]

Animal models

Model organisms have been used in the study of RAD18 function. A conditional knockout mouse line, called Rad18^{tm1a(EUCOMM)Wtsi},^[4] was generated as part of the EUCOMM program — a high-throughput mutagenesis project to generate and distribute animal models of disease to interested scientists — at the Wellcome Trust Sanger Institute.^{[5][6][7][8][9]} Mice lacking Rad18 had no significant defects in viability or fertility,^[10][11] therefore male and female animals underwent a standardized phenotypic screen to determine the effects of deletion.^[6][12][13]

Rad18 knockout mouse phenotype

Characteristic	Phenotype
Homozygote viability	Normal
Fertility	Normal
Body weight	Abnormal[14]
Anxiety	Normal
Neurological assessment	Normal
Grip strength	Normal
Hot plate	Normal
Dysmorphology	Normal[15]
Indirect calorimetry	Abnormal[16]
Glucose tolerance test	Normal
Auditory brainstem response	Normal
DEXA	Abnormal[17]
Radiography	Normal
Body temperature	Normal
Eye morphology	Normal
Clinical chemistry	Normal
Plasma immunoglobulins	Normal
Haematology	Normal
Peripheral blood lymphocytes	Normal
Micronucleus test	Abnormal
Heart weight	Normal
Tail epidermis wholemount	Normal
Skin Histopathology	Normal
Salmonella infection	Normal[18]
Citrobacter infection	Normal[19]
All tests and analysis from[12][20]

Twenty five tests were carried out and four significant phenotypes were reported:^[13]

• Mutant male mice had a decreased body weight compared to wildtype control mice.
• Mutant male mice showed increased activity, VO2 and energy expenditure, determined by indirect calorimetry.
• Dual-energy X-ray absorptiometry (DEXA) showed mutant male mice had a decrease in fat mass.
• A micronucleus test found a potential increase in DNA damage in mutant mice.

Interactions

RAD18 has been shown to interact with HLTF,^[21] UBE2B^[1][2] and UBE2A.^[1][2]

References

1. Tateishi S, Sakuraba Y, Masuyama S, Inoue H, Yamaizumi M (Aug 2000). "Dysfunction of human Rad18 results in defective postreplication repair and hypersensitivity to multiple mutagens". Proc Natl Acad Sci U S A 97 (14): 7927–32. doi:10.1073/pnas.97.14.7927. PMC 16647. PMID 10884424.
2. Xin H, Lin W, Sumanasekera W, Zhang Y, Wu X, Wang Z (Sep 2000). "The human RAD18 gene product interacts with HHR6A and HHR6B". Nucleic Acids Res 28 (14): 2847–54. doi:10.1093/nar/28.14.2847. PMC 102657. PMID 10908344.
3. "Entrez Gene: RAD18 RAD18 homolog (S. cerevisiae)".
4. EUCOMM. "Rad18^{tm1a(EUCOMM)Wtsi}". www.knockoutmouse.org. External link in |publisher= (help)
5. 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.
6. Van der Weyden L, White JK, Adams, DA, Logan DW (June 2011). "The mouse genetics toolkit: revealing function and mechanism". Genome Biology 12 (6): 224. doi:10.1186/gb-2011-12-6-224. PMC 3218837. PMID 21722353.
7. Dolgin E (June 2011). "Mouse library set to be knockout". Nature 474: 262–263. doi:10.1038/474262a. PMID 21677718.
8. Collins FS, Rossant J, Wurst W (January 2007). "A mouse for all reasons". Cell 128 (1): 9–13. doi:10.1016/j.cell.2006.12.018. PMID 17218247.
9. Auwerx J, Avner P, Baldock R, et al. (September 2004). "The European dimension for the mouse genome mutagenesis program". Nat. Genet. 36 (9): 925–7. doi:10.1038/ng0904-925. PMC 2716028. PMID 15340424.
10. Wellcome Trust Sanger Institute. "Viability at Weaning Data for Rad18". Mouse Resources Portal. www.sanger.ac.uk. External link in |publisher= (help)
11. Wellcome Trust Sanger Institute. "Fertility Data for Rad18". Mouse Resources Portal. www.sanger.ac.uk. External link in |publisher= (help)
12. 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.
13. Wellcome Trust Sanger Institute. "MGP Phenotyping of Rad18^{tm1a(EUCOMM)Wtsi}". Mouse Resources Portal. www.sanger.ac.uk. External link in |publisher= (help)
14. "Body weight data for Rad18". Wellcome Trust Sanger Institute.
15. "Dysmorphology data for Rad18". Wellcome Trust Sanger Institute.
16. "Indirect calorimetry data for Rad18". Wellcome Trust Sanger Institute.
17. "DEXA data for Rad18". Wellcome Trust Sanger Institute.
18. "Salmonella infection data for Rad18". Wellcome Trust Sanger Institute.
19. "Citrobacter infection data for Rad18". Wellcome Trust Sanger Institute.
20. Mouse Resources Portal, Wellcome Trust Sanger Institute.
21. Unk, Ildiko; Hajdú Ildikó; Fátyol Károly; Hurwitz Jerard; Yoon Jung-Hoon; Prakash Louise; Prakash Satya; Haracska Lajos (Mar 2008). "Human HLTF functions as a ubiquitin ligase for proliferating cell nuclear antigen polyubiquitination". Proc. Natl. Acad. Sci. U.S.A. (United States) 105 (10): 3768–73. doi:10.1073/pnas.0800563105. PMC 2268824. PMID 18316726.

Further reading

• Maruyama K, Sugano S (1994). "Oligo-capping: a simple method to replace the cap structure of eukaryotic mRNAs with oligoribonucleotides.". Gene 138 (1-2): 171–4. doi:10.1016/0378-1119(94)90802-8. PMID 8125298.
• Suzuki Y, Yoshitomo-Nakagawa K, Maruyama K, et al. (1997). "Construction and characterization of a full length-enriched and a 5'-end-enriched cDNA library.". Gene 200 (1-2): 149–56. doi:10.1016/S0378-1119(97)00411-3. PMID 9373149.
• Mulder LC, Chakrabarti LA, Muesing MA (2002). "Interaction of HIV-1 integrase with DNA repair protein hRad18.". J. Biol. Chem. 277 (30): 27489–93. doi:10.1074/jbc.M203061200. PMID 12016221.
• Strausberg RL, Feingold EA, Grouse LH, et al. (2003). "Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences.". Proc. Natl. Acad. Sci. U.S.A. 99 (26): 16899–903. doi:10.1073/pnas.242603899. PMC 139241. PMID 12477932.
• Nikiforov AA, Sasina LK, Svetlova MP, et al. (2004). "Early immobilization of nuclease FEN1 and accumulation of hRAD18 protein at stalled DNA replication forks in mammalian cells.". Dokl. Biochem. Biophys. 389: 122–5. doi:10.1023/A:1023696425171. PMID 12856420.
• Ota T, Suzuki Y, Nishikawa T, et al. (2004). "Complete sequencing and characterization of 21,243 full-length human cDNAs.". Nat. Genet. 36 (1): 40–5. doi:10.1038/ng1285. PMID 14702039.
• Beausoleil SA, Jedrychowski M, Schwartz D, et al. (2004). "Large-scale characterization of HeLa cell nuclear phosphoproteins.". Proc. Natl. Acad. Sci. U.S.A. 101 (33): 12130–5. doi:10.1073/pnas.0404720101. PMC 514446. PMID 15302935.
• Watanabe K, Tateishi S, Kawasuji M, et al. (2005). "Rad18 guides poleta to replication stalling sites through physical interaction and PCNA monoubiquitination.". EMBO J. 23 (19): 3886–96. doi:10.1038/sj.emboj.7600383. PMC 522788. PMID 15359278.
• Nikiforov A, Svetlova M, Solovjeva L, et al. (2004). "DNA damage-induced accumulation of Rad18 protein at stalled replication forks in mammalian cells involves upstream protein phosphorylation.". Biochem. Biophys. Res. Commun. 323 (3): 831–7. doi:10.1016/j.bbrc.2004.08.165. PMID 15381075.
• Gerhard DS, Wagner L, Feingold EA, et al. (2004). "The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC).". Genome Res. 14 (10B): 2121–7. doi:10.1101/gr.2596504. PMC 528928. PMID 15489334.
• Miyase S, Tateishi S, Watanabe K, et al. (2005). "Differential regulation of Rad18 through Rad6-dependent mono- and polyubiquitination.". J. Biol. Chem. 280 (1): 515–24. doi:10.1074/jbc.M409219200. PMID 15509568.
• Masuyama S, Tateishi S, Yomogida K, et al. (2005). "Regulated expression and dynamic changes in subnuclear localization of mammalian Rad18 under normal and genotoxic conditions.". Genes Cells 10 (8): 753–62. doi:10.1111/j.1365-2443.2005.00874.x. PMID 16098139.
• Nousiainen M, Silljé HH, Sauer G, et al. (2006). "Phosphoproteome analysis of the human mitotic spindle.". Proc. Natl. Acad. Sci. U.S.A. 103 (14): 5391–6. doi:10.1073/pnas.0507066103. PMC 1459365. PMID 16565220.
• Bi X, Barkley LR, Slater DM, et al. (2006). "Rad18 regulates DNA polymerase kappa and is required for recovery from S-phase checkpoint-mediated arrest.". Mol. Cell. Biol. 26 (9): 3527–40. doi:10.1128/MCB.26.9.3527-3540.2006. PMC 1447421. PMID 16611994.
• Lloyd AG, Tateishi S, Bieniasz PD, et al. (2006). "Effect of DNA repair protein Rad18 on viral infection.". PLoS Pathog. 2 (5): e40. doi:10.1371/journal.ppat.0020040. PMC 1463017. PMID 16710452.
• Yuasa MS, Masutani C, Hirano A, et al. (2006). "A human DNA polymerase eta complex containing Rad18, Rad6 and Rev1; proteomic analysis and targeting of the complex to the chromatin-bound fraction of cells undergoing replication fork arrest.". Genes Cells 11 (7): 731–44. doi:10.1111/j.1365-2443.2006.00974.x. PMID 16824193.
• Beausoleil SA, Villén J, Gerber SA, et al. (2006). "A probability-based approach for high-throughput protein phosphorylation analysis and site localization.". Nat. Biotechnol. 24 (10): 1285–92. doi:10.1038/nbt1240. PMID 16964243.
• Olsen JV, Blagoev B, Gnad F, et al. (2006). "Global, in vivo, and site-specific phosphorylation dynamics in signaling networks.". Cell 127 (3): 635–48. doi:10.1016/j.cell.2006.09.026. PMID 17081983.