Reviewed, UniProtKB/Swiss-Prot Q99638 (RAD9A_HUMAN)

Names and origin

Protein names

Recommended name:
    Cell cycle checkpoint control protein RAD9A
      Short name=hRAD9
    EC=3.1.11.2
Alternative name(s):
    DNA repair exonuclease rad9 homolog A

Gene names

Name:

RAD9A

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

General annotation (Comments)

Function	Component of the 9-1-1 cell-cycle checkpoint response complex that plays a major role in DNA repair. The 9-1-1 complex is recruited to DNA lesion upon damage by the RAD17-replication factor C (RFC) clamp loader complex. Acts then as a sliding clamp platform on DNA for several proteins involved in long-patch base excision repair (LP-BER). The 9-1-1 complex stimulates DNA polymerase beta (POLB) activity by increasing its affinity for the 3'-OH end of the primer-template and stabilizes POLB to those sites where LP-BER proceeds; endonuclease FEN1 cleavage activity on substrates with double, nick, or gap flaps of distinct sequences and lengths; and DNA ligase I (LIG1) on long-patch base excision repair substrates. RAD9A possesses 3'->5' double stranded DNA exonuclease activity. Its phosphorylation by PRKCD may be required for the formation of the 9-1-1 complex. Ref.8
Catalytic activity	Exonucleolytic cleavage in the 3'- to 5'-direction to yield nucleoside 5'-phosphates.
Subunit structure	Component of the toroidal 9-1-1 (RAD9-RAD1-HUS1) complex, composed of RAD9A, RAD1 and HUS1. The 9-1-1 complex associates with LIG1, POLB, FEN1, RAD17, HDAC1, RPA1 and RPA2. The 9-1-1 complex associates with the RAD17-RFC complex. RAD9A interacts with BCL2L1, FEN1, PRKCD, RAD9B, HUS1, RAD1, ABL1, RPA1, ATAD5 and RPA2. Ref.6 Ref.7 Ref.11 Ref.12 Ref.13 Ref.18 Ref.19
Subcellular location	Nucleus. Ref.13
Post-translational modification	Constitutively phosphorylated on serine and threonine amino acids in absence of DNA damage. Hyperphosphorylated by PRKCD and ABL1 upon DNA damage. Its phosphorylation by PRKCD may be required for the formation of the 9-1-1 complex. Ref.11 Ref.13 Ref.5 Ref.20 Ref.21
Sequence similarities	Belongs to the rad9 family.

Ontologies

Keywords
Biological process	DNA damage DNA repair
Cellular component	Nucleus
Coding sequence diversity	Polymorphism
Molecular function	Exonuclease Hydrolase Nuclease
PTM	Phosphoprotein
Technical term	Direct protein sequencing
Gene Ontology (GO)
Biological process	DNA damage checkpoint Ref.1 Traceable author statement. Source: ProtInc DNA repair Ref.1 Traceable author statement. Source: ProtInc DNA replication Inferred from Experiment. Source: Reactome DNA replication checkpoint Ref.1 Traceable author statement. Source: ProtInc
Cellular component	nucleus Ref.13 Inferred from direct assay. Source: UniProtKB
Molecular function	3'-5' exonuclease activity Ref.8 Inferred from direct assay. Source: UniProtKB SH3 domain binding Ref.11 Inferred from physical interaction. Source: UniProtKB exodeoxyribonuclease III activity Inferred from electronic annotation. Source: EC histone deacetylase binding Ref.9 Inferred from physical interaction. Source: UniProtKB protein kinase binding Ref.13 Inferred from physical interaction. Source: UniProtKB
Complete GO annotation...

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 391	391	Cell cycle checkpoint control protein RAD9A		PRO_0000225000
Regions
	Region	51 – 91	41	Possesses 3'-5' exonuclease activity
	Region	266 – 391	126	Sufficient for interaction with ABL1
Amino acid modifications
	Modified residue	28	1	Phosphotyrosine Ref.11
	Modified residue	272	1	Phosphoserine Ref.5
	Modified residue	277	1	Phosphoserine Ref.5 Ref.21
	Modified residue	328	1	Phosphoserine Ref.5
	Modified residue	341	1	Phosphoserine Ref.5
	Modified residue	375	1	Phosphoserine Ref.5 Ref.20 Ref.21
	Modified residue	380	1	Phosphoserine Ref.5 Ref.20
	Modified residue	387	1	Phosphoserine Ref.5 Ref.20 Ref.21
Natural variations
	Natural variant	3	1	C → F Ref.3		VAR_025410
	Natural variant	71	1	L → Q: dbSNP rs2422490.		VAR_051724
	Natural variant	100	1	S → A: dbSNP rs2066492. Ref.3		VAR_025411
	Natural variant	239	1	H → R: dbSNP rs17880039. Ref.3		VAR_025412
	Natural variant	307	1	M → T: dbSNP rs17882466. Ref.3		VAR_025413
Experimental info
	Mutagenesis	28	1	Y → F: Abolishes phosphorylation by ABL1. Ref.11
	Mutagenesis	272	1	S → A: Complete loss of phosphorylation and no loss of interaction with the 9-1-1 complex; when associated with A-277; A-328; A-341; A-375; A-380 and A-387. Ref.5
	Mutagenesis	277	1	S → A: Complete loss of phosphorylation and no loss of interaction with the 9-1-1 complex; when associated with A-272; A-328; A-341; A-375; A-380 and A-387. Ref.5
	Mutagenesis	328	1	S → A: Complete loss of phosphorylation and no loss of interaction with the 9-1-1 complex; when associated with A-272; A-277; A-341; A-375; A-380 and A-387. Ref.5
	Mutagenesis	341	1	S → A: Complete loss of phosphorylation and no loss of interaction with the 9-1-1 complex; when associated with A-272; A-277; A-328; A-375; A-380 and A-387. Ref.5
	Mutagenesis	375	1	S → A: Complete loss of phosphorylation and no loss of interaction with the 9-1-1 complex; when associated with A-272; A-277; A-328; A-341; A-380 and A-387. Ref.5
	Mutagenesis	380	1	S → A: Complete loss of phosphorylation and no loss of interaction with the 9-1-1 complex; when associated with A-272; A-277; A-328; A-341; A-375 and A-387. Ref.5
	Mutagenesis	387	1	S → A: Complete loss of phosphorylation and no loss of interaction with the 9-1-1 complex; when associated with A-272; A-277; A-328; A-341; A-375 and A-380. Ref.5
	Sequence conflict	12	1	V → A in CAG38746. Ref.2
	Sequence conflict	130	1	E → A in AAH14848. Ref.4

Sequences

	Sequence		Length	Mass (Da)
	Q99638-1 [UniParc]. Last modified May 1, 1997. Version 1. Checksum: 4D4D6D4C6E1057D3	FASTA	391	42,547
	10 20 30 40 50 60 MKCLVTGGNV KVLGKAVHSL SRIGDELYLE PLEDGLSLRT VNSSRSAYAC FLFAPLFFQQ 70 80 90 100 110 120 YQAATPGQDL LRCKILMKSF LSVFRSLAML EKTVEKCCIS LNGRSSRLVV QLHCKFGVRK 130 140 150 160 170 180 THNLSFQDCE SLQAVFDPAS CPHMLRAPAR VLGEAVLPFS PALAEVTLGI GRGRRVILRS 190 200 210 220 230 240 YHEEEADSTA KAMVTEMCLG EEDFQQLQAQ EGVAITFCLK EFRGLLSFAE SANLNLSIHF 250 260 270 280 290 300 DAPGRPAIFT IKDSLLDGHF VLATLSDTDS HSQDLGSPER HQPVPQLQAH STPHPDDFAN 310 320 330 340 350 360 DDIDSYMIAM ETTIGNEGSR VLPSISLSPG PQPPKSPGPH SEEEDEAEPS TVPGTPPPKK 370 380 390 FRSLFFGSIL APVRSPQGPS PVLAEDSEGE G

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"A human homolog of the Schizosaccharomyces pombe rad9+ checkpoint control gene." Lieberman H.B., Hopkins K.M., Nass M., Demetrick D., Davey S. Proc. Natl. Acad. Sci. U.S.A. 93:13890-13895(1996) [PubMed: 8943031] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA].
[2]	"Cloning of human full open reading frames in Gateway(TM) system entry vector (pDONR201)." Ebert L., Schick M., Neubert P., Schatten R., Henze S., Korn B. Submitted (JUN-2004) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
[3]	NIEHS SNPs program Submitted (SEP-2004) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], VARIANTS PHE-3; ALA-100; ARG-239 AND THR-307.
[4]	"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: Testis.
[5]	"Phosphorylation of human Rad9 is required for genotoxin-activated checkpoint signaling." Roos-Mattjus P., Hopkins K.M., Oestreich A.J., Vroman B.T., Johnson K.L., Naylor S., Lieberman H.B., Karnitz L.M. J. Biol. Chem. 278:24428-24437(2003) [PubMed: 12709442] [Abstract] Cited for: PROTEIN SEQUENCE OF 270-280; 326-329; 373-383 AND 385-390, IDENTIFICATION BY MASS SPECTROMETRY, PHOSPHORYLATION AT SER-272; SER-277; SER-328; SER-341; SER-375; SER-380 AND SER-387, MUTAGENESIS OF SER-272; SER-277; SER-328; SER-341; SER-375; SER-380 AND SER-387.
[6]	"The human G2 checkpoint control protein hRAD9 is a nuclear phosphoprotein that forms complexes with hRAD1 and hHUS1." St Onge R.P., Udell C.M., Casselman R., Davey S. Mol. Biol. Cell 10:1985-1995(1999) [PubMed: 10359610] [Abstract] Cited for: INTERACTION WITH HUS1 AND RAD1.
[7]	"Physical interaction among human checkpoint control proteins HUS1p, RAD1p, and RAD9p, and implications for the regulation of cell cycle progression." Hang H., Lieberman H.B. Genomics 65:24-33(2000) [PubMed: 10777662] [Abstract] Cited for: INTERACTION WITH HUS1 AND RAD1.
[8]	"Human DNA damage checkpoint protein hRAD9 is a 3' to 5' exonuclease." Bessho T., Sancar A. J. Biol. Chem. 275:7451-7454(2000) [PubMed: 10713044] [Abstract] Cited for: FUNCTION IN EXONUCLEASE ACTIVITY.
[9]	"HDAC1, a histone deacetylase, forms a complex with Hus1 and Rad9, two G2/M checkpoint Rad proteins." Cai R.L., Yan-Neale Y., Cueto M.A., Xu H., Cohen D. J. Biol. Chem. 275:27909-27916(2000) [PubMed: 10846170] [Abstract] Cited for: IDENTIFICATION IN THE 9-1-1 COMPLEX ASSOCIATED WITH HDAC1.
[10]	"The human checkpoint protein hRad17 interacts with the PCNA-like proteins hRad1, hHus1, and hRad9." Rauen M., Burtelow M.A., Dufault V.M., Karnitz L.M. J. Biol. Chem. 275:29767-29771(2000) [PubMed: 10884395] [Abstract] Cited for: IDENTIFICATION IN THE 9-1-1 COMPLEX ASSOCIATED WITH RAD17.
[11]	"c-Abl tyrosine kinase regulates the human Rad9 checkpoint protein in response to DNA damage." Yoshida K., Komatsu K., Wang H.-G., Kufe D. Mol. Cell. Biol. 22:3292-3300(2002) [PubMed: 11971963] [Abstract] Cited for: INTERACTION WITH ABL1 AND BCL2L1, PHOSPHORYLATION, PHOSPHORYLATION AT TYR-28, MUTAGENESIS OF TYR-28.
[12]	"Expression of mammalian paralogues of HRAD9 and Mrad9 checkpoint control genes in normal and cancerous testicular tissue." Hopkins K.M., Wang X., Berlin A., Hang H., Thaker H.M., Lieberman H.B. Cancer Res. 63:5291-5298(2003) [PubMed: 14500360] [Abstract] Cited for: INTERACTION WITH RAD9B.
[13]	"Protein kinase Cdelta is responsible for constitutive and DNA damage-induced phosphorylation of Rad9." Yoshida K., Wang H.-G., Miki Y., Kufe D. EMBO J. 22:1431-1441(2003) [PubMed: 12628935] [Abstract] Cited for: INTERACTION WITH PRKCD, PHOSPHORYLATION, SUBCELLULAR LOCATION.
[14]	"Loading of the human 9-1-1 checkpoint complex onto DNA by the checkpoint clamp loader hRad17-replication factor C complex in vitro." Bermudez V.P., Lindsey-Boltz L.A., Cesare A.J., Maniwa Y., Griffith J.D., Hurwitz J., Sancar A. Proc. Natl. Acad. Sci. U.S.A. 100:1633-1638(2003) [PubMed: 12578958] [Abstract] Cited for: ASSOCIATION OF THE 9-1-1 COMPLEX WITH THE RAD17-RFC COMPLEX, ELECTRON MICROSCOPY OF THE 9-1-1 AND RAD17-RFC COMPLEXES BOUND TO DNA.
[15]	"The human Rad9/Rad1/Hus1 damage sensor clamp interacts with DNA polymerase beta and increases its DNA substrate utilisation efficiency: implications for DNA repair." Toueille M., El-Andaloussi N., Frouin I., Freire R., Funk D., Shevelev I., Friedrich-Heineken E., Villani G., Hottiger M.O., Huebscher U. Nucleic Acids Res. 32:3316-3324(2004) [PubMed: 15314187] [Abstract] Cited for: IDENTIFICATION IN THE 9-1-1 COMPLEX ASSOCIATED WITH POLB.
[16]	"The human Rad9-Rad1-Hus1 checkpoint complex stimulates flap endonuclease 1." Wang W., Brandt P., Rossi M.L., Lindsey-Boltz L., Podust V., Fanning E., Sancar A., Bambara R.A. Proc. Natl. Acad. Sci. U.S.A. 101:16762-16767(2004) [PubMed: 15556996] [Abstract] Cited for: IDENTIFICATION IN THE 9-1-1 COMPLEX ASSOCIATED WITH FEN1.
[17]	"The human checkpoint sensor and alternative DNA clamp Rad9-Rad1-Hus1 modulates the activity of DNA ligase I, a component of the long-patch base excision repair machinery." Smirnova E., Toueille M., Markkanen E., Huebscher U. Biochem. J. 389:13-17(2005) [PubMed: 15871698] [Abstract] Cited for: IDENTIFICATION IN THE 9-1-1 COMPLEX ASSOCIATED WITH LIG1.
[18]	"The two DNA clamps Rad9/Rad1/Hus1 complex and proliferating cell nuclear antigen differentially regulate flap endonuclease 1 activity." Friedrich-Heineken E., Toueille M., Taennler B., Buerki C., Ferrari E., Hottiger M.O., Huebscher U. J. Mol. Biol. 353:980-989(2005) [PubMed: 16216273] [Abstract] Cited for: IDENTIFICATION IN THE 9-1-1 COMPLEX ASSOCIATED WITH FEN1, INTERACTION WITH FEN1.
[19]	"Interaction and colocalization of Rad9/Rad1/Hus1 checkpoint complex with replication protein A in human cells." Wu X., Shell S.M., Zou Y. Oncogene 24:4728-4735(2005) [PubMed: 15897895] [Abstract] Cited for: IDENTIFICATION IN THE 9-1-1 COMPLEX ASSOCIATED WITH RPA1 AND RPA2, INTERACTION WITH RPA1 AND RPA2.
[20]	"Global, in vivo, and site-specific phosphorylation dynamics in signaling networks." Olsen J.V., Blagoev B., Gnad F., Macek B., Kumar C., Mortensen P., Mann M. Cell 127:635-648(2006) [PubMed: 17081983] [Abstract] Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-375; SER-380 AND SER-387, MASS SPECTROMETRY. Tissue: Epithelium.
[21]	"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-277; SER-375 AND SER-387, MASS SPECTROMETRY.
+	Additional computationally mapped references.

Web resources

NIEHS-SNPs

Cross-references

Sequence databases
EMBLGenBankDDBJ	U53174 mRNA. Translation: AAB39928.1. CR536508 mRNA. Translation: CAG38746.1. AY766122 Genomic DNA. Translation: AAU89725.1. BC014848 mRNA. Translation: AAH14848.1.
IPI	IPI00005277.
RefSeq	NP_004575.1.
UniGene	Hs.655354
3D structure databases
ModBase	Search...
PTM databases
PhosphoSite	Q99638.
Proteomic databases
PeptideAtlas	Q99638.
PRIDE	Q99638.
Genome annotation databases
Ensembl	ENSG00000172613. Homo sapiens. [Contig view]
GeneID	5883.
KEGG	hsa:5883.
Organism-specific databases
GeneCards	GC11P066915.
H-InvDB	HIX0022833.
HGNC	HGNC:9827. RAD9A.
HPA	HPA006725.
MIM	603761. gene.
PharmGKB	PA294.
GenAtlas	Search...
Phylogenomic databases
HOGENOM	Q99638.
HOVERGEN	Q99638.
OMA	Q99638. SYMIAME.
Enzyme and pathway databases
BRENDA	3.1.11.2. 247.
Pathway_Interaction_DB	telomerasepathway. Regulation of Telomerase.
Reactome	REACT_1538. Cell Cycle Checkpoints.
Gene expression databases
ArrayExpress	Q99638.
Bgee	Q99638.
CleanEx	HS_RAD9A.
GermOnline	ENSG00000172613. Homo sapiens.
Family and domain databases
InterPro	IPR016552. Cell_cycle_RAD9. IPR007268. Rad9. [Graphical view]
PANTHER	PTHR15237. Rad9. 1 hit.
Pfam	PF04139. Rad9. 1 hit. [Graphical view]
PIRSF	PIRSF009303. Cell_cycle_RAD9. 1 hit.
ProtoNet	Search...
Other Resources
NextBio	22862.
SOURCE	Search...

Entry information

Entry name

RAD9A_HUMAN

Accession

Primary (citable) accession number: Q99638
Secondary accession number(s): Q6FI29, Q96C41

Entry history

Integrated into UniProtKB/Swiss-Prot:	March 7, 2006
Last sequence update:	May 1, 1997
Last modified:	June 16, 2009
This is version 52 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HPI (Human Proteome Initiative)