Reviewed, UniProtKB/Swiss-Prot O76064 (RNF8_HUMAN)

Names and origin

Protein names

Recommended name:
    E3 ubiquitin-protein ligase RNF8
    EC=6.3.2.-
Alternative name(s):
    RING finger protein 8

Gene names

Name:	RNF8
Synonyms:	KIAA0646

Organism

Homo sapiens (Human)

Taxonomic identifier

9606 [NCBI]

Taxonomic lineage

Eukaryota › Metazoa › Chordata › Craniata › Vertebrata › Euteleostomi › Mammalia › Eutheria › Euarchontoglires › Primates › Haplorrhini › Catarrhini › Hominidae › Homo

Protein attributes

Sequence length	485 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is not processed.
Protein existence	Evidence at protein level.

General annotation (Comments)

Function	E3 ubiquitin-protein ligase which may be required for proper exit from mitosis after spindle checkpoint activation and may regulate cytokinesis. Promotes the formation of 'Lys-63'-linked polyubiquitin chains and functions with the specific ubiquitin-conjugating UBC13-MMS2 (UBE2N-UBE2V2) heterodimer. Substrates that are polyubiquitinated at 'Lys-63' are usually not targeted for degradation. Following DNA double-strand breaks (DSB), is recruited to the sites of damage by ATM-phosphorylated MDC1 and promotes the formation of TP53BP1 and BRCA1 ionizing radiation-induced foci (IRIF). Responsible for 'Lys-63'-linked ubiquitin chains at DSB sites. Enforces the G2/M DNA damage checkpoint. May play a role in the regulation of RXRA-mediated transcriptional activity. Not involved in RXRA ubiquitination by UBE2E2. Ref.9 Ref.10 Ref.11 Ref.12
Pathway	Protein modification; protein ubiquitination.
Subunit structure	Interacts with class III E2s, including UBE2E1, UBE2E2, and UBE2E3 and with UBE2N. Interacts with RXRA. Interacts with ATM-phosphorylated MDC1. Ref.9 Ref.10 Ref.11 Ref.2
Subcellular location	Nucleus. Note= 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. Following DNA double-strand breaks, recruited to the sites of damage. Ref.9 Ref.10 Ref.2
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. Ref.2 Ref.1
Developmental stage	Low levels at the G1-S boundary increase in intensity during S phase and until the end of the G2 phase. Abruptly decreases in late mitosis (at protein level). Barely detectable in anaphase. Ref.12
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.
Sequence similarities	Contains 1 FHA domain. Contains 1 RING-type zinc finger.

Ontologies

Keywords
Biological process	Cell cycle Cell division DNA damage Mitosis Ubl conjugation pathway
Cellular component	Nucleus
Coding sequence diversity	Polymorphism
Domain	Zinc-finger
Ligand	Metal-binding Zinc
Molecular function	Ligase
PTM	Phosphoprotein Ubl conjugation
Technical term	3D-structure
Gene Ontology (GO)
Biological process	cell division Inferred from electronic annotation. Source: UniProtKB-KW mitosis Inferred from electronic annotation. Source: UniProtKB-KW response to DNA damage stimulus Inferred from electronic annotation. Source: UniProtKB-KW ubiquitin-dependent protein catabolic process Inferred from electronic annotation. Source: UniProtKB-KW
Cellular component	nucleus Inferred from electronic annotation. Source: UniProtKB-KW
Molecular function	ligase activity Inferred from electronic annotation. Source: UniProtKB-KW protein binding Inferred from physical interaction. Source: IntAct zinc ion binding Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
EXOSC2	Q13868	1	EBI-373337,EBI-301735
MDC1	Q14676	5	EBI-373337,EBI-495644
ORC2L	Q13416	1	EBI-373337,EBI-374957

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 485

485

E3 ubiquitin-protein ligase RNF8

PRO_0000056048

Regions

Domain

38 – 92

55

FHA

Zinc finger

403 – 441

39

RING-type

Compositional bias

276 – 345

70

Gln-rich

Amino acid modifications

Modified residue

157

1

Phosphoserine Ref.13

Natural variations

Natural variant

162

1

A → T: dbSNP rs34338974.

VAR_052096

Natural variant

473

1

I → V: dbSNP rs1139944.

VAR_052097

Experimental info

Mutagenesis

403

1

C → S: Marked reduction of E2-dependent ubiquitination. Loss of UBE2E2- and UBE2N-binding. Loss of nuclear localization Ref.9 Ref.10 Ref.2

Sequence conflict

230

1

V → A in BAD96485. Ref.5

Sequence conflict

334

1

K → R in BAD96485. Ref.5

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

485

Helix Strand Turn

Details...

Beta strand

15 – 20

6

Beta strand

29 – 31

3

Beta strand

38 – 43

6

Beta strand

46 – 49

4

Helix

56 – 58

3

Beta strand

64 – 68

5

Beta strand

74 – 80

7

Beta strand

85 – 89

5

Beta strand

106 – 109

4

Beta strand

124 – 128

5

Helix

129 – 132

4

Sequences

	Sequence		Length	Mass (Da)
	O76064-1 [UniParc]. Last modified November 1, 1998. Version 1. Checksum: 54650B2FFC9948B1	FASTA	485	55,518
	10 20 30 40 50 60 MGEPGFFVTG DRAGGRSWCL RRVGMSAGWL LLEDGCEVTV GRGFGVTYQL VSKICPLMIS 70 80 90 100 110 120 RNHCVLKQNP EGQWTIMDNK SLNGVWLNRA RLEPLRVYSI HQGDYIQLGV PLENKENAEY 130 140 150 160 170 180 EYEVTEEDWE TIYPCLSPKN DQMIEKNKEL RTKRKFSLDE LAGPGAEGPS NLKSKINKVS 190 200 210 220 230 240 CESGQPVKSQ GKGEVASTPS DNLDPKLTAL EPSKTTGAPI YPGFPKVTEV HHEQKASNSS 250 260 270 280 290 300 ASQRSLQMFK VTMSRILRLK IQMQEKHEAV MNVKKQTQKG NSKKVVQMEQ ELQDLQSQLC 310 320 330 340 350 360 AEQAQQQARV EQLEKTFQEE EQHLQGLEIA QGEKDLKQQL AQALQEHWAL MEELNRSKKD 370 380 390 400 410 420 FEAIIQAKNK ELEQTKEEKE KMQAQKEEVL SHMNDVLENE LQCIICSEYF IEAVTLNCAH 430 440 450 460 470 480 SFCSYCINEW MKRKIECPIC RKDIKSKTYS LVLDNCINKM VNNLSSEVKE RRIVLIRERK AKRLF

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Isolation, tissue expression, and chromosomal assignment of a novel human gene which encodes a protein with RING finger motif." Seki N., Hattori A., Sugano S., Suzuki Y., Nakagawara A., Ohhira M., Muramatsu M., Hori T., Saito T. J. Hum. Genet. 43:272-274(1998) [PubMed: 9852682] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], TISSUE SPECIFICITY. Tissue: Brain.
[2]	"The RING finger protein RNF8 recruits UBC13 for lysine 63-based self polyubiquitylation." Plans V., Scheper J., Soler M., Loukili N., Okano Y., Thomson T.M. J. Cell. Biochem. 97:572-582(2006) [PubMed: 16215985] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], SUBCELLULAR LOCATION, TISSUE SPECIFICITY, INTERACTION WITH UBE2E2 AND UBE2N, AUTOUBIQUITINATION, MUTAGENESIS OF CYS-403. Tissue: Fetal brain.
[3]	"Prediction of the coding sequences of unidentified human genes. X. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro." Ishikawa K., Nagase T., Suyama M., Miyajima N., Tanaka A., Kotani H., Nomura N., Ohara O. DNA Res. 5:169-176(1998) [PubMed: 9734811] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA]. Tissue: Brain.
[4]	"Cloning of human full-length CDSs in BD Creator(TM) system donor vector." Kalnine N., Chen X., Rolfs A., Halleck A., Hines L., Eisenstein S., Koundinya M., Raphael J., Moreira D., Kelley T., LaBaer J., Lin Y., Phelan M., Farmer A. Submitted (MAY-2003) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
[5]	Suzuki Y., Sugano S., Totoki Y., Toyoda A., Takeda T., Sakaki Y., Tanaka A., Yokoyama S. Submitted (APR-2005) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA]. Tissue: Hepatoma.
[6]	"The DNA sequence and analysis of human chromosome 6." Mungall A.J., Palmer S.A., Sims S.K., Edwards C.A., Ashurst J.L., Wilming L., Jones M.C., Horton R., Hunt S.E., Scott C.E., Gilbert J.G.R., Clamp M.E., Bethel G., Milne S., Ainscough R., Almeida J.P., Ambrose K.D., Andrews T.D. , Ashwell R.I.S., Babbage A.K., Bagguley C.L., Bailey J., Banerjee R., Barker D.J., Barlow K.F., Bates K., Beare D.M., Beasley H., Beasley O., Bird C.P., Blakey S.E., Bray-Allen S., Brook J., Brown A.J., Brown J.Y., Burford D.C., Burrill W., Burton J., Carder C., Carter N.P., Chapman J.C., Clark S.Y., Clark G., Clee C.M., Clegg S., Cobley V., Collier R.E., Collins J.E., Colman L.K., Corby N.R., Coville G.J., Culley K.M., Dhami P., Davies J., Dunn M., Earthrowl M.E., Ellington A.E., Evans K.A., Faulkner L., Francis M.D., Frankish A., Frankland J., French L., Garner P., Garnett J., Ghori M.J., Gilby L.M., Gillson C.J., Glithero R.J., Grafham D.V., Grant M., Gribble S., Griffiths C., Griffiths M.N.D., Hall R., Halls K.S., Hammond S., Harley J.L., Hart E.A., Heath P.D., Heathcott R., Holmes S.J., Howden P.J., Howe K.L., Howell G.R., Huckle E., Humphray S.J., Humphries M.D., Hunt A.R., Johnson C.M., Joy A.A., Kay M., Keenan S.J., Kimberley A.M., King A., Laird G.K., Langford C., Lawlor S., Leongamornlert D.A., Leversha M., Lloyd C.R., Lloyd D.M., Loveland J.E., Lovell J., Martin S., Mashreghi-Mohammadi M., Maslen G.L., Matthews L., McCann O.T., McLaren S.J., McLay K., McMurray A., Moore M.J.F., Mullikin J.C., Niblett D., Nickerson T., Novik K.L., Oliver K., Overton-Larty E.K., Parker A., Patel R., Pearce A.V., Peck A.I., Phillimore B.J.C.T., Phillips S., Plumb R.W., Porter K.M., Ramsey Y., Ranby S.A., Rice C.M., Ross M.T., Searle S.M., Sehra H.K., Sheridan E., Skuce C.D., Smith S., Smith M., Spraggon L., Squares S.L., Steward C.A., Sycamore N., Tamlyn-Hall G., Tester J., Theaker A.J., Thomas D.W., Thorpe A., Tracey A., Tromans A., Tubby B., Wall M., Wallis J.M., West A.P., White S.S., Whitehead S.L., Whittaker H., Wild A., Willey D.J., Wilmer T.E., Wood J.M., Wray P.W., Wyatt J.C., Young L., Younger R.M., Bentley D.R., Coulson A., Durbin R.M., Hubbard T., Sulston J.E., Dunham I., Rogers J., Beck S. Nature 425:805-811(2003) [PubMed: 14574404] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
[7]	Mural R.J., Istrail S., Sutton G.G., Florea L., Halpern A.L., Mobarry C.M., Lippert R., Walenz B., Shatkay H., Dew I., Miller J.R., Flanigan M.J., Edwards N.J., Bolanos R., Fasulo D., Halldorsson B.V., Hannenhalli S., Turner R. , Yooseph S., Lu F., Nusskern D.R., Shue B.C., Zheng X.H., Zhong F., Delcher A.L., Huson D.H., Kravitz S.A., Mouchard L., Reinert K., Remington K.A., Clark A.G., Waterman M.S., Eichler E.E., Adams M.D., Hunkapiller M.W., Myers E.W., Venter J.C. Submitted (JUL-2005) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
[8]	"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: 15489334] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA]. Tissue: Muscle.
[9]	"N-terminally extended human ubiquitin-conjugating enzymes (E2s) mediate the ubiquitination of RING-finger proteins, ARA54 and RNF8." Ito K., Adachi S., Iwakami R., Yasuda H., Muto Y., Seki N., Okano Y. Eur. J. Biochem. 268:2725-2732(2001) [PubMed: 11322894] [Abstract] Cited for: FUNCTION, SUBCELLULAR LOCATION, INTERACTION WITH UBE2E1; UBE2E2 AND UBE2E3, MUTAGENESIS OF CYS-403.
[10]	"The RING finger protein, RNF8, interacts with retinoid X receptor alpha and enhances its transcription-stimulating activity." Takano Y., Adachi S., Okuno M., Muto Y., Yoshioka T., Matsushima-Nishiwaki R., Tsurumi H., Ito K., Friedman S.L., Moriwaki H., Kojima S., Okano Y. J. Biol. Chem. 279:18926-18934(2004) [PubMed: 14981089] [Abstract] Cited for: FUNCTION, SUBCELLULAR LOCATION, INTERACTION WITH RXRA, MUTAGENESIS OF CYS-403.
[11]	"Orchestration of the DNA-damage response by the RNF8 ubiquitin ligase." Kolas N.K., Chapman J.R., Nakada S., Ylanko J., Chahwan R., Sweeney F.D., Panier S., Mendez M., Wildenhain J., Thomson T.M., Pelletier L., Jackson S.P., Durocher D. Science 318:1637-1640(2007) [PubMed: 18006705] [Abstract] Cited for: FUNCTION, INTERACTION WITH MDC1.
[12]	"Regulation of mitotic exit by the RNF8 ubiquitin ligase." Plans V., Guerra-Rebollo M., Thomson T.M. Oncogene 27:1355-1365(2008) [PubMed: 17724460] [Abstract] Cited for: FUNCTION, DEVELOPMENTAL STAGE.
[13]	"A quantitative atlas of mitotic phosphorylation." Dephoure N., Zhou C., Villen J., Beausoleil S.A., Bakalarski C.E., Elledge S.J., Gygi S.P. Proc. Natl. Acad. Sci. U.S.A. 105:10762-10767(2008) [PubMed: 18669648] [Abstract] Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-157, MASS SPECTROMETRY.
[14]	"Solution structure of the FHA domain of human ubiquitin ligase protein RNF8." RIKEN structural genomics initiative (RSGI) Submitted (NOV-2005) to the PDB data bank Cited for: STRUCTURE BY NMR OF 8-139.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBLGenBankDDBJ

AF334675 mRNA. Translation: AAQ14887.1.
AB012770 Genomic DNA. Translation: BAA33557.1.
AB014546 mRNA. Translation: BAA31621.2. Different initiation.
BT007446 mRNA. Translation: AAP36114.1.
AK222765 mRNA. Translation: BAD96485.1.
AL096712 Genomic DNA. Translation: CAB75689.1.
CH471081 Genomic DNA. Translation: EAX03944.1.
BC007517 mRNA. Translation: AAH07517.1.

RefSeq

NP_003949.1.
NP_898901.1.

UniGene

Hs.485278

3D structure databases

PDBMSD

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
2CSW	NMR	-	A	8-139	[»]
2PIE	X-ray	1.35	A	13-146	[»]

SMR

O76064. Positions 8-140.

ModBase

Search...

Protein-protein interaction databases

IntAct

O76064. 13 interactions.

PTM databases

PhosphoSite

O76064.

Proteomic databases

PRIDE

O76064.

Genome annotation databases

Ensembl

ENSG00000112130. Homo sapiens. [Contig view]

GeneID

9025.

KEGG

hsa:9025.

Organism-specific databases

GeneCards

GC06P037429.

H-InvDB

HIX0005839.

HGNC

HGNC:10071. RNF8.

MIM

611685. gene.

PharmGKB

PA34445.

HUGE

Search...

GenAtlas

Search...

Phylogenomic databases

HOGENOM

O76064.

HOVERGEN

O76064.

Gene expression databases

ArrayExpress

O76064.

CleanEx

HS_RNF8.

GermOnline

ENSG00000112130. Homo sapiens.

Family and domain databases

InterPro

IPR017335. E3_ubiquit_lig_RNF8.
IPR000253. FHA.
IPR001841. Znf_RING.
IPR013083. Znf_RING/FYVE/PHD.
[Graphical view]

Gene3D

G3DSA:2.60.200.20. FHA. 1 hit.
G3DSA:3.30.40.10. Znf_RING/FYVE/PHD. 1 hit.

Pfam

PF00498. FHA. 1 hit.
PF00097. zf-C3HC4. 1 hit.
[Graphical view]

PIRSF

PIRSF037950. E3_ubiquit_lig_RNF8. 1 hit.

SMART

SM00240. FHA. 1 hit.
SM00184. RING. 1 hit.
[Graphical view]

PROSITE

PS50006. FHA_DOMAIN. 1 hit.
PS00518. ZF_RING_1. 1 hit.
PS50089. ZF_RING_2. 1 hit.
[Graphical view]

ProtoNet

Search...

Other Resources

NextBio

33813.

SOURCE

Search...

Entry information

Entry name

RNF8_HUMAN

Accession

Primary (citable) accession number: O76064
Secondary accession number(s): Q53H16, Q5NKW5

Entry history

Integrated into UniProtKB/Swiss-Prot:	June 6, 2002
Last sequence update:	November 1, 1998
Last modified:	December 16, 2008
This is version 85 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HPI (Human Proteome Initiative)