Recombinant Human E3 Ubiquitin-Protein Ligase Rnf8 (RNF8) Protein (His&Myc)

Beta LifeScience SKU/CAT #: BLC-01427P
Greater than 85% as determined by SDS-PAGE.
Greater than 85% as determined by SDS-PAGE.

Recombinant Human E3 Ubiquitin-Protein Ligase Rnf8 (RNF8) Protein (His&Myc)

Beta LifeScience SKU/CAT #: BLC-01427P
Regular price $936.00 Sale price $349.00Save $587
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Product Overview

Description Recombinant Human E3 Ubiquitin-Protein Ligase Rnf8 (RNF8) Protein (His&Myc) is produced by our Baculovirus expression system. This is a full length protein.
Purity Greater than 85% as determined by SDS-PAGE.
Uniprotkb O76064
Target Symbol RNF8
Synonyms RING finger protein 8 (RING-type E3 ubiquitin transferase RNF8)
Species Homo sapiens (Human)
Expression System Baculovirus
Tag N-10His&C-Myc
Target Protein Sequence MGEPGFFVTGDRAGGRSWCLRRVGMSAGWLLLEDGCEVTVGRGFGVTYQLVSKICPLMISRNHCVLKQNPEGQWTIMDNKSLNGVWLNRARLEPLRVYSIHQGDYIQLGVPLENKENAEYEYEVTEEDWETIYPCLSPKNDQMIEKNKELRTKRKFSLDELAGPGAEGPSNLKSKINKVSCESGQPVKSQGKGEVASTPSDNLDPKLTALEPSKTTGAPIYPGFPKVTEVHHEQKASNSSASQRSLQMFKVTMSRILRLKIQMQEKHEAVMNVKKQTQKGNSKKVVQMEQELQDLQSQLCAEQAQQQARVEQLEKTFQEEEQHLQGLEIAQGEKDLKQQLAQALQEHWALMEELNRSKKDFEAIIQAKNKELEQTKEEKEKMQAQKEEVLSHMNDVLENELQSIISSEYFIEAVTLNCAHSFCSYCINEWMKRKIECPICRKDIKSKTYSLVLDNCINKMVNNLSSEVKERRIVLIRERKAKRLF
Expression Range 1-485aa(C403S, C406S)
Protein Length Full Length
Mol. Weight 59.4 kDa
Research Area Cell Biology
Form Liquid or Lyophilized powder
Buffer Liquid form: default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol. Lyophilized powder form: the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.
Reconstitution Briefly centrifuged the vial prior to opening to bring the contents to the bottom. Reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL. It is recommended to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. The default final concentration of glycerol is 50%.
Storage 1. Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. 2. Avoid repeated freeze-thaw cycles. 3. Store working aliquots at 4°C for up to one week. 4. In general, protein in liquid form is stable for up to 6 months at -20°C/-80°C. Protein in lyophilized powder form is stable for up to 12 months at -20°C/-80°C.
Notes Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Target Details

Target Function E3 ubiquitin-protein ligase that plays a key role in DNA damage signaling via 2 distinct roles: by mediating the 'Lys-63'-linked ubiquitination of histones H2A and H2AX and promoting the recruitment of DNA repair proteins at double-strand breaks (DSBs) sites, and by catalyzing 'Lys-48'-linked ubiquitination to remove target proteins from DNA damage sites. Following DNA DSBs, it is recruited to the sites of damage by ATM-phosphorylated MDC1 and catalyzes the 'Lys-63'-linked ubiquitination of histones H2A and H2AX, thereby promoting the formation of TP53BP1 and BRCA1 ionizing radiation-induced foci (IRIF). Also controls the recruitment of UIMC1-BRCC3 (RAP80-BRCC36) and PAXIP1/PTIP to DNA damage sites. Also recruited at DNA interstrand cross-links (ICLs) sites and catalyzes 'Lys-63'-linked ubiquitination of histones H2A and H2AX, leading to recruitment of FAAP20/C1orf86 and Fanconi anemia (FA) complex, followed by interstrand cross-link repair. H2A ubiquitination also mediates the ATM-dependent transcriptional silencing at regions flanking DSBs in cis, a mechanism to avoid collision between transcription and repair intermediates. Promotes the formation of 'Lys-63'-linked polyubiquitin chains via interactions with the specific ubiquitin-conjugating UBE2N/UBC13 and ubiquitinates non-histone substrates such as PCNA. Substrates that are polyubiquitinated at 'Lys-63' are usually not targeted for degradation. Also catalyzes the formation of 'Lys-48'-linked polyubiquitin chains via interaction with the ubiquitin-conjugating UBE2L6/UBCH8, leading to degradation of substrate proteins such as CHEK2, JMJD2A/KDM4A and KU80/XRCC5: it is still unclear how the preference toward 'Lys-48'- versus 'Lys-63'-linked ubiquitination is regulated but it could be due to RNF8 ability to interact with specific E2 specific ligases. For instance, interaction with phosphorylated HERC2 promotes the association between RNF8 and UBE2N/UBC13 and favors the specific formation of 'Lys-63'-linked ubiquitin chains. Promotes non-homologous end joining (NHEJ) by promoting the 'Lys-48'-linked ubiquitination and degradation the of KU80/XRCC5. Following DNA damage, mediates the ubiquitination and degradation of JMJD2A/KDM4A in collaboration with RNF168, leading to unmask H4K20me2 mark and promote the recruitment of TP53BP1 at DNA damage sites. Following DNA damage, mediates the ubiquitination and degradation of POLD4/p12, a subunit of DNA polymerase delta. In the absence of POLD4, DNA polymerase delta complex exhibits higher proofreading activity. In addition to its function in damage signaling, also plays a role in higher-order chromatin structure by mediating extensive chromatin decondensation. Involved in the activation of ATM by promoting histone H2B ubiquitination, which indirectly triggers histone H4 'Lys-16' acetylation (H4K16ac), establishing a chromatin environment that promotes efficient activation of ATM kinase. Required in the testis, where it plays a role in the replacement of histones during spermatogenesis. At uncapped telomeres, promotes the joining of deprotected chromosome ends by inducing H2A ubiquitination and TP53BP1 recruitment, suggesting that it may enhance cancer development by aggravating telomere-induced genome instability in case of telomeric crisis. Promotes the assembly of RAD51 at DNA DSBs in the absence of BRCA1 and TP53BP1 Also involved in class switch recombination in immune system, via its role in regulation of DSBs repair. May be required for proper exit from mitosis after spindle checkpoint activation and may regulate cytokinesis. May play a role in the regulation of RXRA-mediated transcriptional activity. Not involved in RXRA ubiquitination by UBE2E2.
Subcellular Location Nucleus. Cytoplasm. Midbody. Chromosome, telomere.
Protein Families RNF8 family
Database References
Tissue Specificity Ubiquitous. In fetal tissues, highest expression in brain, thymus and liver. In adult tissues, highest levels in brain and testis, lowest levels in peripheral blood cells.

Gene Functions References

  1. study describes RNF8 as a co-activator of ERalpha increases ERalpha stability via post-transcriptional pathway, and provides a new insight into mechanisms for RNF8 to promote cell growth of ERalpha-positive breast cancer. PMID: 28216286
  2. the present study described the noncovalent interaction between the E3 ubiquitin ligase RNF8 and SUMO2/3 and indicated that this interaction promoted DSB repair. PMID: 28983621
  3. Data report that RNF8 is overexpressed in highly metastatic breast cell lines and its overexpression can induce EMT in breast cancer cells. Furthermore, RNF8 is aberrantly expressed in invasive breast cancer and positively correlates with lymph node metastasis. PMID: 27259701
  4. The present findings indicate that WRAP53beta and RNF8 are rate-limiting factors in the repair of DNA double-strand breaks and raise the possibility that upregulation of WRAP53beta may contribute to genomic stability in and survival of cancer cells. PMID: 27310875
  5. The study identifies a previously unrecognized role for RNF8 in the negative regulation of NF-kappaB activation by targeting and deactivating the IKK complex. PMID: 28499869
  6. the role of RNF8-mediated histone H3 polyubiquitylation in the regulation of histone H3 stability and chromatin modification, is reported. PMID: 28507061
  7. RNF8- and Ube2S-dependent Lys11-linkage ubiquitin conjugation plays an important role in regulating DNA damage-induced transcriptional silencing, distinct from the role of Lys63-linkage ubiquitin in the recruitment of DNA damage repair proteins 53BP1 and BRCA1. PMID: 28525740
  8. RNF8-promoted Twist ubiquitination is required for Twist localization to the nucleus for subsequent epithelial-mesenchymal transition and cancer stem cells functions, thereby conferring chemoresistance. PMID: 27618486
  9. The attenuated DNA damage localization of RNF8 resulting from INT6 depletion could be attributed to the defective retention of ATM. PMID: 27550454
  10. High RNF8 expression is associated with bladder cancer. PMID: 26788910
  11. The interaction of MDC1 with RNF8, but not with ATM requires WRAP53beta, suggesting that WRAP53beta facilitates the former interaction without altering phosphorylation of MDC1 by ATM. PMID: 26734725
  12. define a novel function for ATDC in the RNF8-mediated DNA damage response and implicate RNF8 binding as a key determinant of the radioprotective function of ATDC PMID: 26381412
  13. Results indicate that RNF8 recruits and ubiquitinates many factors to repair DNA damage thereby conferring radioresistance to nasopharyngeal cancer cells. PMID: 25955491
  14. Significantly restored tolerance of RAD18-/- and RNF8-/- cells to camptothecin and olaparib without affecting Rad51 focus formation. PMID: 25417706
  15. considered potential associations of 14 single nucleotide polymorphisms (SNPs) in RNF8 and BRDT genes in Chinese patients with non-obstructive azoospermia PMID: 25374327
  16. our results indicate that down-regulation of RNF8 mediated by miR-214 impedes DNA damage response to induce chromosomal instability in ovarian cancers, which may facilitate the understanding of mechanisms underlying chromosomal instability. PMID: 25483088
  17. Finding that RNF8 is less abundant than RNF168 identifies RNF8 as a rate-limiting determinant of focal repair complex assembly. PMID: 25304081
  18. Depletion of RNF8 or RNF168 blocks the degradation of diffusely localized nuclear 53BP1. PMID: 25337968
  19. PALB2 localization depends on the presence of MDC1, RNF8, RAP80 and Abraxas upstream of BRCA1. PMID: 23038782
  20. RNF8 maintains genome stability through independent, yet analogous, nuclear and cytoplasmic ubiquitylation activities PMID: 23442799
  21. findings implicate USP44 in negative regulation of the RNF8/RNF168 pathway PMID: 23615962
  22. The PARP1-dependent localization of BAL1-BBAP functionally limits both early and delayed DNA damage and enhances cellular viability independent of ATM, MDC1, and RNF8. PMID: 23230272
  23. The identification of RNF8 allows new insights into the integration of the control of p12 degradation by different DNA damage signaling pathways. PMID: 23233665
  24. Nbs1 is one important target of RNF8 to regulate DNA DSB repair. PMID: 23115235
  25. Data indicate that RNF8-dependent chromatin ubiquitination is required for RAD51 assembly in BRCA1/53BP1-depleted cells. PMID: 22865450
  26. RNF8 may play a role in protein synthesis, possibly linking the nucleolar exit of this factor to the attenuation of protein synthesis in response to DNA damage. PMID: 22814251
  27. Data indicate that RNF8 and FAAP20 (C1orf86) are needed for efficient Fanconi anemia group D2 protein FANCD2 monoubiquitination. PMID: 22705371
  28. A new mechanism of chromatin remodelling-assisted ubiquitylation was shown, which involves cooperation between CHD4 and RNF8 to create a local chromatin environment permissive to the assembly of checkpoint and repair machineries at DNA lesions. PMID: 22531782
  29. The authors propose that the RNF8-dependent degradation of JMJD2A regulates DNA repair by controlling the recruitment of 53BP1 at DNA damage sites. PMID: 22373579
  30. By mimicking a cellular phosphosite, ICP0 binds RNF8 via the RNF8 forkhead associated (FHA) domain. Phosphorylation of ICP0 T67 by CK1 recruits RNF8 for degradation and thereby promotes viral transcription PMID: 22405594
  31. Data show that depletion of RNF8, as well as of the E3 ligase RNF168, reduces telomere-induced genome instability. PMID: 21857671
  32. The viral protein ICP0 targets RNF8 and RNF168 for degradation, thereby preventing the deposition of repressive ubiquitin marks and counteracting this repair protein recruitment. PMID: 21698222
  33. the differential requirement for the ubiquitin ligase RNF8 in facilitating repair of replication stress-associated DNA damage PMID: 21558560
  34. The E3 Ubiquitin ligases, RNF8 and RNF168, are recruited to DNA damage foci in late mitosis, presumably to prime sites for the DNA damage response, 53BP1, recruitment in early G1. PMID: 21412056
  35. These results suggest that RNF8 is downregulated in many cancer cells and inversely correlated with Plk1. PMID: 21635870
  36. Phosphorylated NPM1 may interact with RNF8-dependent ubiquitin conjugates at sites of DNA damage. PMID: 20713529
  37. Data identify RNF8 and RNF168, cellular histone ubiquitin ligases responsible for anchoring repair factors at sites of damage, as new targets for ICP0-mediated degradation. PMID: 20075863
  38. Ubiquitin ligase does not protect cells from Nutlin-3-mediated apoptosis, indicating that RNF8 does not regulate 53BP1 protein. PMID: 20080757
  39. Data show that the ATM signalling mediator proteins MDC1, RNF8, RNF168 and 53BP1 are also required for heterochromatic DSB repair. PMID: 20081839
  40. results suggest a novel function of RNF8 as a regulator of RXR alpha-mediated transcriptional activity through interaction between their respective N-terminal regions PMID: 14981089
  41. Regulates the rate of exit from mitosis and cytokinesis. PMID: 17724460
  42. RNF8 ubiquitylates histones at DNA double-strand breaks and promotes assembly of repair proteins. PMID: 18001824
  43. RNF8 is a novel DNA-damage-responsive protein that integrates protein phosphorylation and ubiquitylation signaling and plays a critical role in the cellular response to genotoxic stress. PMID: 18001825
  44. results demonstrate how the DNA-damage response is orchestrated by ATM-dependent phosphorylation of MDC1 and RNF8-mediated ubiquitination PMID: 18006705
  45. RNF8 is the human orthologue of the yeast protein Dma1p PMID: 18171988
  46. RNF8 ubiquitylation pathways are essential for 53BP1 regulation in response to ionizing radiation, whereas RNF8-independent pathways contribute to 53BP1 targeting and phosphorylation in response to UV light and other forms of DNA replication stress PMID: 18337245
  47. Depletion of RAP80 or RNF8 impairs the translocation of BRCA1 to DNA damage sites and results in defective cell cycle checkpoint control and DSB repair PMID: 18550271
  48. RNF8 readily mono-ubiquitinates PCNA in the presence of UbcH5c, and polyubiquitinates PCNA in the added presence of Ubc13/Uev1a. PMID: 18948756
  49. subset of PTIP.PA1 complex is recruited to DNA damage sites via the RNF8-dependent pathway and is required for cell survival in response to DNA damage. PMID: 19124460
  50. Nucleotide excision repair-induced H2A ubiquitination is dependent on MDC1 and RNF8 and reveals a universal DNA damage response. PMID: 19797077

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