Q8VC56 (RNF8_MOUSE) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
E3 ubiquitin-protein ligase RNF8
EC=6.3.2.-

Alternative name(s):
ActA-interacting protein 37
Short name=AIP37
LaXp180
RING finger protein 8

Gene names

Name:

Rnf8

Organism

Mus musculus (Mouse) [Reference proteome]

Taxonomic identifier

10090 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Opisthokonta › Metazoa › Eumetazoa › Bilateria › Deuterostomia › Chordata › Craniata › Vertebrata › Gnathostomata › Teleostomi › Euteleostomi › Sarcopterygii › Tetrapoda › Amniota › Mammalia › Theria › Eutheria › Euarchontoglires › Glires › Rodentia › Sciurognathi › Muroidea › Muridae › Murinae › Mus › Mus

Protein attributes

Sequence length	488 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

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 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. 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 aggraving 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. Ref.5 Ref.6 Ref.7 Ref.8 Ref.10
Pathway	Protein modification; protein ubiquitination.
Subunit structure	Homodimer. Forms a E2-E3 ubiquitin ligase complex composed of the RNF8 homodimer and a E2 heterodimer of UBE2N and UBE2V2. Interacts with class III E2s, including UBE2E1, UBE2E2, and UBE2E3 and with UBE2N. Interacts with RXRA. Interacts (via FHA domain) with ATM-phosphorylated MDC1. Interacts (via FHA domain) with 'Thr-4829' phosphorylated HERC2 (via C-terminus) By similarity. May interact with the L.monocytogenes protein actA; however, given these errors in the sequence (AJ242721), the relevance of the interation with actA remains to be confirmed. Ref.4
Subcellular location	Nucleus By similarity. Midbody By similarity. Chromosome › telomere. Note: Following DNA double-strand breaks, recruited to the sites of damage. During prophase, concomitant with nuclear envelope breakdown, localizes throughout the cell, with a dotted pattern. In telophase, again in the nucleus and also with a discrete dotted pattern in the cytoplasm. In late telophase and during cytokinesis, localizes in the midbody of the tubulin bridge joining the daughter cells. Does not seem to be associated with condensed chromosomes at any time during the cell cycle By similarity. Recruited at uncapped telomeres. Ref.8 Ref.9
Domain	The FHA domain specifically recognizes and binds ATM-phosphorylated MDC1 and 'Thr-4829' phosphorylated HERC2 By similarity.
Post-translational modification	Autoubiquitinated through 'Lys-48' and 'Lys-63' of ubiquitin. 'Lys-63' polyubiquitination is mediated by UBE2N. 'Lys-29'-type polyubiquitination is also observed, but it doesn't require its own functional RING-type zinc finger By similarity.
Disruption phenotype	Male mice are infertile, while females do not show defects. Male mice display defects in histone H2A and H2B ubiquitination in testis cells. While meiotic sex chromosome inactivation in the XY body prior to meiosis is not affected, H4K16ac is decreased, leading to defects in the replacement of histones by protamines during spermiogenesis. Mice lacking both Rnf8 and Chfr develop thymic lymphomas and chromosomes are frequently altered, due to defects in DNA damage response and defects in damage-induced activation of ATM kinase. Ref.5 Ref.7
Sequence similarities	Belongs to the RNF8 family. Contains 1 FHA domain. Contains 1 RING-type zinc finger.
Caution	The precise role of Rnf8 at telomeres is subject to debate. 2 publications reported recruitment of Rnf8 at uncapped telomeres followed by regulation of non-homologous end joining (NHEJ), however the 2 publications reported different data and conclusions. According to a report, Rnf8 promotes telomere end protection and inhibits NHEJ by mediating ubiquitination via 'Lys-63'-linked ubiquitin and stabilization of Tpp1 at uncapped telomeres (Ref.9). According to another report, Rnf8 recruitment at uncapped telomeres leads to promote NHEJ and the joining of deprotected chromosome ends by inducing H2A ubiquitination and TP53BP1 recruitment, suggesting that Rnf8 may have a detrimental role in case of telomeric crisis and enhance cancer development by aggraving telomere-induced genome instability (Ref.8). According to a well-established model, RNF8 initiate H2A 'Lys-63'-linked ubiquitination leading to recruitment of RNF168 to amplify H2A 'Lys-63'-linked ubiquitination. However, other data suggest that RNF168 is the priming ubiquitin ligase by mediating monoubiquitination of 'Lys-13' and 'Lys-15' of nucleosomal histone H2A (H2AK13Ub and H2AK15Ub respectively). These data suggest that RNF168 might be recruited to DSBs sites in a RNF8-dependent manner by binding to non-histone proteins ubiquitinated via 'Lys-63'-linked and initiates monoubiquitination of H2A, which is then amplified by RNF8. Additional evidences are however required to confirm these data.
Sequence caution	The sequence CAB92239.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally shortened. The sequence CAB92239.1 differs from that shown. Reason: Frameshift at positions 219, 266 and 283.

Ontologies

Keywords
Biological process	Cell cycle Cell division DNA damage DNA repair Mitosis Ubl conjugation pathway
Cellular component	Chromosome Nucleus Telomere
Domain	Zinc-finger
Ligand	Metal-binding Zinc
Molecular function	Chromatin regulator Ligase
PTM	Phosphoprotein Ubl conjugation
Technical term	Complete proteome Reference proteome
Gene Ontology (GO)
Biological_process	cell division Inferred from electronic annotation. Source: UniProtKB-KW double-strand break repair via nonhomologous end joining Inferred from mutant phenotype Ref.8Ref.10. Source: UniProtKB histone H2A K63-linked ubiquitination Inferred from sequence or structural similarity. Source: UniProtKB histone H2B ubiquitination Inferred from mutant phenotype PubMed 19015238Ref.5. Source: UniProtKB histone exchange Inferred from mutant phenotype Ref.5. Source: UniProtKB isotype switching Inferred from mutant phenotype Ref.6. Source: UniProtKB mitosis Inferred from electronic annotation. Source: UniProtKB-KW negative regulation of translational elongation Inferred from sequence or structural similarity. Source: UniProtKB positive regulation of DNA repair Inferred from sequence or structural similarity. Source: UniProtKB protein K48-linked ubiquitination Inferred from sequence or structural similarity. Source: UniProtKB protein autoubiquitination Inferred from electronic annotation. Source: Compara response to ionizing radiation Inferred from sequence or structural similarity. Source: UniProtKB spermatid development Inferred from mutant phenotype Ref.5. Source: UniProtKB ubiquitin-dependent protein catabolic process Inferred from sequence or structural similarity. Source: UniProtKB
Cellular_component	chromosome, telomeric region Inferred from direct assay Ref.8. Source: UniProtKB midbody Inferred from electronic annotation. Source: UniProtKB-SubCell nucleus Inferred from sequence or structural similarity. Source: UniProtKB site of double-strand break Inferred from sequence or structural similarity. Source: UniProtKB ubiquitin ligase complex Inferred from sequence or structural similarity. Source: UniProtKB
Molecular_function	chromatin binding Inferred from sequence or structural similarity. Source: UniProtKB histone binding Inferred from mutant phenotype PubMed 19015238. Source: UniProtKB protein homodimerization activity Inferred from sequence or structural similarity. Source: UniProtKB ubiquitin-protein ligase activity Inferred from mutant phenotype PubMed 19015238. Source: UniProtKB zinc ion binding Inferred from sequence or structural similarity. Source: UniProtKB
Complete GO annotation...

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 488	488	E3 ubiquitin-protein ligase RNF8		PRO_0000056049
Regions
	Domain	38 – 92	55	FHA
	Zinc finger	406 – 444	39	RING-type
	Compositional bias	299 – 351	53	Gln-rich
Amino acid modifications
	Modified residue	157	1	Phosphoserine By similarity

Sequences

	Sequence		Length	Mass (Da)
	Q8VC56 [UniParc]. Last modified March 1, 2002. Version 1. Checksum: 428242204EBC44A1	FASTA	488	55,517
	10 20 30 40 50 60 MGEPDPLVSG QLAARRSWCL RRLGMDCEWL QLEAGTEVTI GRGLSVTYQL ISKVCPLMIS 70 80 90 100 110 120 RSHCVLKQNP EGQWTIMDNK SLNGVWLNRE RLAPLQGYCI RKGDHIQLGV PLESRETAEY 130 140 150 160 170 180 EYEVIEEDWE SLAPCLAPKN DQRMEKHKGS RTKRKFSSPG LENLPAEGSS DLRCPLANVA 190 200 210 220 230 240 SKPIEPEKLH GKGDASSQSL GCLCPGLTSL KASERAAGPH ACSALPKVLE LSCPKKQKAC 250 260 270 280 290 300 RPSASQNSLE LFKVTMSRML KLKTQMQEKQ IAVLNVKRQT RKGSSKKIVR MEKELRNLQS 310 320 330 340 350 360 QLYAEQAQQQ ARVEQLEKTF QEEAHYLQGL EKEQGECDLK QQLVQALQEH QALMEELNCS 370 380 390 400 410 420 KKDFEKIIQA KNKELEQTKE EKDKVQAQKE EVLSHMNDLL ENELQCIICS EYFIEAVTLN 430 440 450 460 470 480 CAHSFCSFCI NEWMKRKVEC PICRKDIESR TNSLVLDNCI SKMVDNLSSD VKERRSVLIR ERRAKRLS

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"The transcriptional landscape of the mammalian genome." Carninci P., Kasukawa T., Katayama S., Gough J., Frith M.C., Maeda N., Oyama R., Ravasi T., Lenhard B., Wells C., Kodzius R., Shimokawa K., Bajic V.B., Brenner S.E., Batalov S., Forrest A.R., Zavolan M., Davis M.J. , Wilming L.G., Aidinis V., Allen J.E., Ambesi-Impiombato A., Apweiler R., Aturaliya R.N., Bailey T.L., Bansal M., Baxter L., Beisel K.W., Bersano T., Bono H., Chalk A.M., Chiu K.P., Choudhary V., Christoffels A., Clutterbuck D.R., Crowe M.L., Dalla E., Dalrymple B.P., de Bono B., Della Gatta G., di Bernardo D., Down T., Engstrom P., Fagiolini M., Faulkner G., Fletcher C.F., Fukushima T., Furuno M., Futaki S., Gariboldi M., Georgii-Hemming P., Gingeras T.R., Gojobori T., Green R.E., Gustincich S., Harbers M., Hayashi Y., Hensch T.K., Hirokawa N., Hill D., Huminiecki L., Iacono M., Ikeo K., Iwama A., Ishikawa T., Jakt M., Kanapin A., Katoh M., Kawasawa Y., Kelso J., Kitamura H., Kitano H., Kollias G., Krishnan S.P., Kruger A., Kummerfeld S.K., Kurochkin I.V., Lareau L.F., Lazarevic D., Lipovich L., Liu J., Liuni S., McWilliam S., Madan Babu M., Madera M., Marchionni L., Matsuda H., Matsuzawa S., Miki H., Mignone F., Miyake S., Morris K., Mottagui-Tabar S., Mulder N., Nakano N., Nakauchi H., Ng P., Nilsson R., Nishiguchi S., Nishikawa S., Nori F., Ohara O., Okazaki Y., Orlando V., Pang K.C., Pavan W.J., Pavesi G., Pesole G., Petrovsky N., Piazza S., Reed J., Reid J.F., Ring B.Z., Ringwald M., Rost B., Ruan Y., Salzberg S.L., Sandelin A., Schneider C., Schoenbach C., Sekiguchi K., Semple C.A., Seno S., Sessa L., Sheng Y., Shibata Y., Shimada H., Shimada K., Silva D., Sinclair B., Sperling S., Stupka E., Sugiura K., Sultana R., Takenaka Y., Taki K., Tammoja K., Tan S.L., Tang S., Taylor M.S., Tegner J., Teichmann S.A., Ueda H.R., van Nimwegen E., Verardo R., Wei C.L., Yagi K., Yamanishi H., Zabarovsky E., Zhu S., Zimmer A., Hide W., Bult C., Grimmond S.M., Teasdale R.D., Liu E.T., Brusic V., Quackenbush J., Wahlestedt C., Mattick J.S., Hume D.A., Kai C., Sasaki D., Tomaru Y., Fukuda S., Kanamori-Katayama M., Suzuki M., Aoki J., Arakawa T., Iida J., Imamura K., Itoh M., Kato T., Kawaji H., Kawagashira N., Kawashima T., Kojima M., Kondo S., Konno H., Nakano K., Ninomiya N., Nishio T., Okada M., Plessy C., Shibata K., Shiraki T., Suzuki S., Tagami M., Waki K., Watahiki A., Okamura-Oho Y., Suzuki H., Kawai J., Hayashizaki Y. Science 309:1559-1563(2005) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA]. Strain: C57BL/6J. Tissue: Extraembryonic tissue, Liver and Placenta.
[2]	"Cloning of mouse full open reading frames in Gateway(R) system entry vector (pDONR201)." Ebert L., Muenstermann E., Schatten R., Henze S., Bohn E., Mollenhauer J., Wiemann S., Schick M., Korn B. Submitted (JUL-2005) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
[3]	"The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC)." The MGC Project Team Genome Res. 14:2121-2127(2004) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA]. Tissue: Kidney.
[4]	"LaXp180, a mammalian ActA-binding protein, identified with the yeast two-hybrid system co-localizes with intracellular Listeria monocytogenes." Pfeuffer T., Goebel W., Laubinger J., Bachmann M., Kuhn M. Cell. Microbiol. 2:101-114(2000) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA] OF 1-283, INTERACTION WITH ACTA. Strain: CD-1. Tissue: Embryo.
[5]	"RNF8-dependent histone modifications regulate nucleosome removal during spermatogenesis." Lu L.Y., Wu J., Ye L., Gavrilina G.B., Saunders T.L., Yu X. Dev. Cell 18:371-384(2010) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, DISRUPTION PHENOTYPE.
[6]	"The RNF8/RNF168 ubiquitin ligase cascade facilitates class switch recombination." Ramachandran S., Chahwan R., Nepal R.M., Frieder D., Panier S., Roa S., Zaheen A., Durocher D., Scharff M.D., Martin A. Proc. Natl. Acad. Sci. U.S.A. 107:809-814(2010) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[7]	"Chfr and RNF8 synergistically regulate ATM activation." Wu J., Chen Y., Lu L.Y., Wu Y., Paulsen M.T., Ljungman M., Ferguson D.O., Yu X. Nat. Struct. Mol. Biol. 18:761-768(2011) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, DISRUPTION PHENOTYPE.
[8]	"DNA-damage response and repair activities at uncapped telomeres depend on RNF8." Peuscher M.H., Jacobs J.J. Nat. Cell Biol. 13:1139-1145(2011) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, SUBCELLULAR LOCATION.
[9]	"The E3 ubiquitin ligase Rnf8 stabilizes Tpp1 to promote telomere end protection." Rai R., Li J.M., Zheng H., Lok G.T., Deng Y., Huen M.S., Chen J., Jin J., Chang S. Nat. Struct. Mol. Biol. 18:1400-1407(2011) [PubMed] [Europe PMC] [Abstract] Cited for: SUBCELLULAR LOCATION.
[10]	"The E3 ligase RNF8 regulates KU80 removal and NHEJ repair." Feng L., Chen J. Nat. Struct. Mol. Biol. 19:201-206(2012) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
+	Additional computationally mapped references.

Cross-references

Sequence databases
EMBL GenBank DDBJ	AK076180 mRNA. Translation: BAC36236.1. AK147168 mRNA. Translation: BAE27732.1. CT010295 mRNA. Translation: CAJ18503.1. BC021778 mRNA. Translation: AAH21778.1. AJ242721 mRNA. Translation: CAB92239.1. Frameshift.
IPI	IPI00551150.
RefSeq	NP_067394.1. NM_021419.1.
UniGene	Mm.305994.
3D structure databases
ProteinModelPortal	Q8VC56.
SMR	Q8VC56. Positions 8-140, 354-486.
ModBase	Search...
Protein-protein interaction databases
DIP	DIP-59448N.
PTM databases
PhosphoSite	Q8VC56.
Proteomic databases
PRIDE	Q8VC56.
Protocols and materials databases
StructuralBiologyKnowledgebase	Search...
Genome annotation databases
Ensembl	ENSMUST00000024817; ENSMUSP00000024817; ENSMUSG00000090083.
GeneID	58230.
KEGG	mmu:58230.
UCSC	uc008btf.1. mouse.
Organism-specific databases
CTD	9025.
MGI	MGI:1929069. Rnf8.
Phylogenomic databases
GeneTree	ENSGT00400000022349.
HOGENOM	HOG000154169.
HOVERGEN	HBG023954.
KO	K10667.
Enzyme and pathway databases
UniPathway	UPA00143.
Gene expression databases
ArrayExpress	Q8VC56.
Bgee	Q8VC56.
CleanEx	MM_RNF8.
Genevestigator	Q8VC56.
GermOnline	ENSMUSG00000024019. Mus musculus.
Family and domain databases
Gene3D	2.60.200.20. 1 hit. 3.30.40.10. 1 hit.
InterPro	IPR017335. E3_Ub_ligase_RNF8. IPR000253. FHA_dom. IPR008984. SMAD_FHA_domain. IPR001841. Znf_RING. IPR013083. Znf_RING/FYVE/PHD. IPR017907. Znf_RING_CS. [Graphical view]
PANTHER	PTHR15067:SF3. PTHR15067:SF3. 1 hit.
Pfam	PF00498. FHA. 1 hit. [Graphical view]
PIRSF	PIRSF037950. E3_ubiquit_lig_RNF8. 1 hit.
SMART	SM00240. FHA. 1 hit. SM00184. RING. 1 hit. [Graphical view]
SUPFAM	SSF49879. SMAD_FHA. 1 hit.
PROSITE	PS50006. FHA_DOMAIN. 1 hit. PS00518. ZF_RING_1. 1 hit. PS50089. ZF_RING_2. 1 hit. [Graphical view]
ProtoNet	Search...
Other
NextBio	314253.
SOURCE	Search...

Entry information

Entry name

RNF8_MOUSE

Accession

Primary (citable) accession number: Q8VC56
Secondary accession number(s): Q4FJV7, Q9JK13

Entry history

Integrated into UniProtKB/Swiss-Prot:	June 6, 2002
Last sequence update:	March 1, 2002
Last modified:	May 1, 2013
This is version 96 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Chordata Protein Annotation Program