AC MF_03067; DC Protein; auto TR HAMAP; MF_03067; -; 1; level=0 XX Names: RNF8 XX ID RNF8 case DE RecName: Full=E3 ubiquitin-protein ligase RNF8; DE EC=2.3.2.27; DE AltName: Full=RING finger protein 8; DE AltName: Full=RING-type E3 ubiquitin transferase RNF8; else DE RecName: Full=E3 ubiquitin-protein ligase ; DE EC=2.3.2.27; DE AltName: Full=RING finger protein 8; DE AltName: Full=RING-type E3 ubiquitin transferase ; end case GN Name=RNF8; XX case CC -!- FUNCTION: E3 ubiquitin-protein ligase that plays a key role in DNA CC damage signaling via 2 distinct roles: by mediating the 'Lys-63'-linked CC ubiquitination of histones H2A and H2AX and promoting the recruitment CC of DNA repair proteins at double-strand breaks (DSBs) sites, and by CC catalyzing 'Lys-48'-linked ubiquitination to remove target proteins CC from DNA damage sites. Following DNA DSBs, it is recruited to the sites CC of damage by ATM-phosphorylated MDC1 and catalyzes the 'Lys-63'-linked CC ubiquitination of histones H2A and H2AX, thereby promoting the CC formation of TP53BP1 and BRCA1 ionizing radiation-induced foci (IRIF). CC Also controls the recruitment of UIMC1-BRCC3 (RAP80-BRCC36) and CC PAXIP1/PTIP to DNA damage sites. Also recruited at DNA interstrand CC cross-links (ICLs) sites and catalyzes 'Lys-63'-linked ubiquitination CC of histones H2A and H2AX, leading to recruitment of FAAP20 and Fanconi CC anemia (FA) complex, followed by interstrand cross-link repair. H2A CC ubiquitination also mediates the ATM-dependent transcriptional CC silencing at regions flanking DSBs in cis, a mechanism to avoid CC collision between transcription and repair intermediates. Promotes the CC formation of 'Lys-63'-linked polyubiquitin chains via interactions with CC the specific ubiquitin-conjugating UBE2N/UBC13 and ubiquitinates non- CC histone substrates such as PCNA. Substrates that are polyubiquitinated CC at 'Lys-63' are usually not targeted for degradation. Also catalyzes CC the formation of 'Lys-48'-linked polyubiquitin chains via interaction CC with the ubiquitin-conjugating UBE2L6/UBCH8, leading to degradation of CC substrate proteins such as CHEK2, JMJD2A/KDM4A and KU80/XRCC5: it is CC still unclear how the preference toward 'Lys-48'- versus 'Lys-63'- CC linked ubiquitination is regulated but it could be due to RNF8 ability CC to interact with specific E2 specific ligases. For instance, CC interaction with phosphorylated HERC2 promotes the association between CC RNF8 and UBE2N/UBC13 and favors the specific formation of 'Lys-63'- CC linked ubiquitin chains. Promotes non-homologous end joining (NHEJ) by CC promoting the 'Lys-48'-linked ubiquitination and degradation the of CC KU80/XRCC5. Following DNA damage, mediates the ubiquitination and CC degradation of JMJD2A/KDM4A in collaboration with RNF168, leading to CC unmask H4K20me2 mark and promote the recruitment of TP53BP1 at DNA CC damage sites. Following DNA damage, mediates the ubiquitination and CC degradation of POLD4/p12, a subunit of DNA polymerase delta. In the CC absence of POLD4, DNA polymerase delta complex exhibits higher CC proofreading activity. In addition to its function in damage signaling, CC also plays a role in higher-order chromatin structure by mediating CC extensive chromatin decondensation. Involved in the activation of ATM CC by promoting histone H2B ubiquitination, which indirectly triggers CC histone H4 'Lys-16' acetylation (H4K16ac), establishing a chromatin CC environment that promotes efficient activation of ATM kinase. Required CC in the testis, where it plays a role in the replacement of histones CC during spermatogenesis. At uncapped telomeres, promotes the joining of CC deprotected chromosome ends by inducing H2A ubiquitination and TP53BP1 CC recruitment, suggesting that it may enhance cancer development by CC aggravating telomere-induced genome instability in case of telomeric CC crisis. Promotes the assembly of RAD51 at DNA DSBs in the absence of CC BRCA1 and TP53BP1 Also involved in class switch recombination in immune CC system, via its role in regulation of DSBs repair. May be required for CC proper exit from mitosis after spindle checkpoint activation and may CC regulate cytokinesis. May play a role in the regulation of RXRA- CC mediated transcriptional activity. Not involved in RXRA ubiquitination CC by UBE2E2. CC -!- SUBUNIT: Homodimer. Forms a E2-E3 ubiquitin ligase complex composed of CC the RNF8 homodimer and a E2 heterodimer of UBE2N and UBE2V2. Interacts CC with class III E2s, including UBE2E1, UBE2E2, and UBE2E3 and with CC UBE2N. Interacts with RXRA. Interacts (via FHA domain) with CC phosphorylated HERC2 (via C-terminus). Interacts with PIWIL1; leading CC to sequester RNF8 in the cytoplasm. CC -!- SUBCELLULAR LOCATION: Nucleus. Cytoplasm. Midbody. Chromosome, CC telomere. Note=Recruited at uncapped telomeres. Following DNA double- CC strand breaks, recruited to the sites of damage. During prophase, CC concomitant with nuclear envelope breakdown, localizes throughout the CC cell, with a dotted pattern. In telophase, again in the nucleus and CC also with a discrete dotted pattern in the cytoplasm. In late telophase CC and during cytokinesis, localizes in the midbody of the tubulin bridge CC joining the daughter cells. Does not seem to be associated with CC condensed chromosomes at any time during the cell cycle. During CC spermatogenesis, sequestered in the cytoplasm by PIWIL1: RNF8 is CC released following ubiquitination and degradation of PIWIL1. CC -!- DOMAIN: The FHA domain specifically recognizes and binds ATM- CC phosphorylated MDC1 and phosphorylated HERC2. else CC -!- FUNCTION: E3 ubiquitin-protein ligase that plays a key role in DNA CC damage signaling via 2 distinct roles: by mediating the 'Lys-63'-linked CC ubiquitination of histones H2A and H2AX and promoting the recruitment CC of DNA repair proteins at double-strand breaks (DSBs) sites, and by CC catalyzing 'Lys-48'-linked ubiquitination to remove target proteins CC from DNA damage sites. Following DNA DSBs, it is recruited to the sites CC of damage by ATM-phosphorylated @gn(MDC1) and catalyzes the 'Lys-63'- CC linked ubiquitination of histones H2A and H2AX, thereby promoting the CC formation of @gn(TP53BP1) and @gn(BRCA1) ionizing radiation-induced CC foci (IRIF). H2A ubiquitination also mediates the ATM-dependent CC transcriptional silencing at regions flanking DSBs in cis, a mechanism CC to avoid collision between transcription and repair intermediates. Also CC catalyzes the formation of 'Lys-48'-linked polyubiquitin chains, CC leading to degradation of substrate proteins. In addition to its CC function in damage signaling, also plays a role in higher-order CC chromatin structure by mediating extensive chromatin decondensation. CC -!- SUBUNIT: Homodimer. Forms a E2-E3 ubiquitin ligase complex composed of CC the @gn(RNF8) homodimer and a E2 heterodimer of @gn(UBE2N) and CC @gn(UBE2V2). CC -!- SUBCELLULAR LOCATION: Nucleus. Note=Following DNA double-strand breaks, CC recruited to the sites of damage. CC -!- DOMAIN: The FHA domain specifically recognizes and binds ATM- CC phosphorylated MDC1. end case CC -!- CATALYTIC ACTIVITY: CC Reaction=S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine + CC [acceptor protein]-L-lysine = [E2 ubiquitin-conjugating enzyme]-L- CC cysteine + N(6)-ubiquitinyl-[acceptor protein]-L-lysine.; CC EC=2.3.2.27; CC -!- PATHWAY: Protein modification; protein ubiquitination. case CC -!- PTM: Autoubiquitinated through 'Lys-48' and 'Lys-63' of ubiquitin. CC 'Lys-63' polyubiquitination is mediated by UBE2N. 'Lys-29'-type CC polyubiquitination is also observed, but it doesn't require its own CC functional RING-type zinc finger. CC -!- CAUTION: According to a well-established model, RNF8 initiate H2A 'Lys- CC 63'-linked ubiquitination leading to recruitment of RNF168 to amplify CC H2A 'Lys-63'-linked ubiquitination. However, other data suggest that CC RNF168 is the priming ubiquitin ligase by mediating monoubiquitination CC of 'Lys-13' and 'Lys-15' of nucleosomal histone H2A (H2AK13Ub and CC H2AK15Ub respectively). These data suggest that RNF168 might be CC recruited to DSBs sites in a RNF8-dependent manner by binding to non- CC histone proteins ubiquitinated via 'Lys-63'-linked and initiates CC monoubiquitination of H2A, which is then amplified by RNF8. Additional CC evidences are however required to confirm these data. end case CC -!- SIMILARITY: Belongs to the RNF8 family. XX DR PROSITE; PS50006; FHA_DOMAIN; 1; trigger=yes DR PROSITE; PS00518; ZF_RING_1; 1; trigger=no DR PROSITE; PS50089; ZF_RING_2; 1; trigger=yes DR PROSITE; PS50322; GLN_RICH; 0-unlimited; trigger=strict DR Pfam; PF00498; FHA; 1; trigger=no XX case KW Cell cycle KW Cell division KW Chromosome KW Cytoplasm KW Mitosis KW Telomere KW Ubl conjugation end case KW Chromatin regulator KW DNA damage KW DNA repair KW Metal-binding KW Nucleus KW Transferase KW Ubl conjugation pathway KW Zinc KW Zinc-finger XX GO GO:0003682; F:chromatin binding GO GO:0042393; F:histone binding GO GO:0043130; F:ubiquitin binding GO GO:0004842; F:ubiquitin-protein transferase activity GO GO:0045739; P:positive regulation of DNA repair GO GO:0010212; P:response to ionizing radiation GO GO:0006302; P:double-strand break repair GO GO:0033522; P:histone H2A ubiquitination GO GO:0033523; P:histone H2B ubiquitination GO GO:0000151; C:ubiquitin ligase complex GO GO:0005634; C:nucleus XX FT From: RNF8_MOUSE (Q8VC56) case FT REGION 68..72 FT /note="Required for interaction with PIWIL1" end case XX Size: 457-550; Related: None; Template: O76064; Q8VC56; Scope: Eukaryota; Vertebrata Fusion: Nter: None Cter: None Duplicate: in XENLA Plasmid: None Comments: None XX # Revision 1.6 2019/11/20 //