Replication protein A 32 kDa subunit (RPA2) Recombinant Protein | RPA2 recombinant protein

Recombinant Human Replication protein A 32 kDa subunit (RPA2), partial

Cat. Num. AAA717206

• Gene Names: RPA2; REPA2; RPA32; RP-A p32; RP-A p34
• Purity: Greater or equal to 85% purity as determined by SDS-PAGE.
• Synonyms: Replication protein A 32 kDa subunit (RPA2); Recombinant Human Replication protein A 32 kDa subunit (RPA2); partial; Replication factor A protein 2; RF-A protein 2; Replication protein A 34 kDa subunit; RP-A p34; RPA2 recombinant protein

• Host: E Coli or Yeast or Baculovirus or Mammalian Cell
• Purity/Purification: Greater or equal to 85% purity as determined by SDS-PAGE.
• Form/Format: Lyophilized or liquid (Format to be determined during the manufacturing process)
• Sequence Positions: 1-267aa; Partial
• Sequence: MWNSGFESYGSSSYGGAGGYTQSPGGFGSPAPSQAEKKSRARAQHIVPCTISQLLSATLVDEVFRIGNVEISQVTIVGIIRHAEKAPTNIVYKIDDMTAAPMDVRQWVDTDDTSSENTVVPPETYVKVAGHLRSFQNKKSLVAFKIMPLEDMNEFTTHILEVINAHMVLSKANSQPSAGRAPISNPGMSEAGNFGGNSFMPANGLTVAQNQVLNLIKACPRPEGLNFQDLKNQLKHMSVSSIKQAVDFLSNEGHIYSTVDDDHFKST

• Preparation and Storage: Store at -20 degrees C. For long-term storage, store at -20 degrees C or -80 degrees C. Store working aliquots at 4 degrees C for up to one week. Repeated freezing and thawing is not recommended.

SDS-PAGE

Related Product Information for RPA2 recombinant protein

As part of the heterotrimeric replication protein A complex (RPA/RP-A), binds and stabilizes single-stranded DNA intermediates, that form during DNA replication or upon DNA stress. It prevents their reannealing and in parallel, recruits and activates different proteins and complexes involved in DNA metabolism. Thereby, it plays an essential role both in DNA replication and the cellular response to DNA damage. In the cellular response to DNA damage, the RPA complex controls DNA repair and DNA damage checkpoint activation. Through recruitment of ATRIP activates the ATR kinase a master regulator of the DNA damage response. It is required for the recruitment of the DNA double-strand break repair factors RAD51 and RAD52 to chromatin in response to DNA damage. Also recruits to sites of DNA damage proteins like XPA and XPG that are involved in nucleotide excision repair and is required for this mechanism of DNA repair. Plays also a role in base excision repair (BER) probably through interaction with UNG. Through RFWD3 may activate CHEK1 and play a role in replication checkpoint control. Also recruits SMARCAL1/HARP, which is involved in replication fork restart, to sites of DNA damage. May also play a role in telomere maintenance

Product Categories/Family for RPA2 recombinant protein

Epigenetics and Nuclear Signaling

References

The primary structure of the 32-kDa subunit of human replication protein A.Erdile L.F., Wold M.S., Kelly T.J.J. Biol. Chem. 265:3177-3182(1990) Cell-cycle-regulated phosphorylation of DNA replication factor A from human and yeast cells.Din S., Brill S.J., Fairman M.P., Stillman B.Genes Dev. 4:968-977(1990) cdc2 family kinases phosphorylate a human cell DNA replication factor, RPA, and activate DNA replication.Dutta A., Stillman B.EMBO J. 11:2189-2199(1992) The ionizing radiation-induced replication protein A phosphorylation response differs between ataxia telangiectasia and normal human cells.Liu V.F., Weaver D.T.Mol. Cell. Biol. 13:7222-7231(1993) Mammalian DNA nucleotide excision repair reconstituted with purified protein components.Aboussekhra A., Biggerstaff M., Shivji M.K., Vilpo J.A., Moncollin V., Podust V.N., Protic M., Huebscher U., Egly J.M., Wood R.D.Cell 80:859-868(1995) RPA involvement in the damage-recognition and incision steps of nucleotide excision repair.He Z., Henricksen L.A., Wold M.S., Ingles C.J.Nature 374:566-569(1995) Physical interaction between human RAD52 and RPA is required for homologous recombination in mammalian cells.Park M.S., Ludwig D.L., Stigger E., Lee S.H.J. Biol. Chem. 271:18996-19000(1996) Mapping of amino acid residues in the p34 subunit of human single-stranded DNA-binding protein phosphorylated by DNA-dependent protein kinase and Cdc2 kinase in vitro.Niu H., Erdjument-Bromage H., Pan Z.-Q., Lee S.-H., Tempst P., Hurwitz J.J. Biol. Chem. 272:12634-12641(1997) Sites of UV-induced phosphorylation of the p34 subunit of replication protein A from HeLa cells.Zernik-Kobak M., Vasunia K., Connelly M., Anderson C.W., Dixon K.J. Biol. Chem. 272:23896-23904(1997) The evolutionarily conserved zinc finger motif in the largest subunit of human replication protein A is required for DNA replication and mismatch repair but not for nucleotide excision repair.Lin Y.L., Shivji M.K., Chen C., Kolodner R., Wood R.D., Dutta A.J. Biol. Chem. 273:1453-1461(1998) Replication protein A stimulates long patch DNA base excision repair.De;Mott M.S., Zigman S., Bambara R.A.J. Biol. Chem. 273:27492-27498(1998) RBT1, a novel transcriptional co-activator, binds the second subunit of replication protein A.Cho J.M., Song D.J., Bergeron J., Benlimame N., Wold M.S., Alaoui-Jamali M.A.Nucleic Acids Res. 28:3478-3485(2000) ATR kinase activity regulates the intranuclear translocation of ATR and RPA following ionizing radiation.Barr S.M., Leung C.G., Chang E.E., Cimprich K.A.Curr. Biol. 13:1047-1051(2003) Coordinated regulation of replication protein A activities by its subunits p14 and p32.Weisshart K., Pestryakov P., Smith R.W.P., Hartmann H., Kremmer E., Lavrik O., Nasheuer H.-P.J. Biol. Chem. 279:35368-35376(2004) Mammalian TIMELESS and Tipin are evolutionarily conserved replication fork-associated factors.Gotter A.L., Suppa C., Emanuel B.S.J. Mol. Biol. 366:36-52(2007) RPA mediates recombination repair during replication stress and is displaced from DNA by checkpoint signalling in human cells.Sleeth K.M., Sorensen C.S., Issaeva N., Dziegielewski J., Bartek J., Helleday T.J. Mol. Biol. 373:38-47(2007) The human Tim/Tipin complex coordinates an Intra-S checkpoint response to UV that slows replication fork displacement.Uensal-Kacmaz K., Chastain P.D., Qu P.-P., Minoo P., Cordeiro-Stone M., Sancar A., Kaufmann W.K.Mol. Cell. Biol. 27:3131-3142(2007) Replication protein A prevents accumulation of single-stranded telomeric DNA in cells that use alternative lengthening of telomeres.Grudic A., Jul-Larsen A., Haring S.J., Wold M.S., Loenning P.E., Bjerkvig R., Boee S.O.Nucleic Acids Res. 35:7267-7278(2007) 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) Lys-N and trypsin cover complementary parts of the phosphoproteome in a refined SCX-based approach.Gauci S., Helbig A.O., Slijper M., Krijgsveld J., Heck A.J., Mohammed S.Anal. Chem. 81:4493-4501(2009) The annealing helicase HARP is recruited to DNA repair sites via an interaction with RPA.Yusufzai T., Kong X., Yokomori K., Kadonaga J.T.Genes Dev. 23:2400-2404(2009) The annealing helicase SMARCAL1 maintains genome integrity at stalled replication forks.Bansbach C.E., Betous R., Lovejoy C.A., Glick G.G., Cortez D.Genes Dev. 23:2405-2414(2009) The SIOD disorder protein SMARCAL1 is an RPA-interacting protein involved in replication fork restart.Ciccia A., Bredemeyer A.L., Sowa M.E., Terret M.E., Jallepalli P.V., Harper J.W., Elledge S.J.Genes Dev. 23:2415-2425(2009) An alternative form of replication protein a prevents viral replication in vitro.Mason A.C., Haring S.J., Pryor J.M., Staloch C.A., Gan T.F., Wold M.S.J. Biol. Chem. 284:5324-5331(2009) An alternative form of replication protein a expressed in normal human tissues supports DNA repair.Kemp M.G., Mason A.C., Carreira A., Reardon J.T., Haring S.J., Borgstahl G.E., Kowalczykowski S.C., Sancar A., Wold M.S.J. Biol. Chem. 285:4788-4797(2010) A PP4 phosphatase complex dephosphorylates RPA2 to facilitate DNA repair via homologous recombination.Lee D.H., Pan Y., Kanner S., Sung P., Borowiec J.A., Chowdhury D.Nat. Struct. Mol. Biol. 17:365-372(2010) Quantitative phosphoproteomics reveals widespread full phosphorylation site occupancy during mitosis.Olsen J.V., Vermeulen M., Santamaria A., Kumar C., Miller M.L., Jensen L.J., Gnad F., Cox J., Jensen T.S., Nigg E.A., Brunak S., Mann M.Sci. Signal. 3:RA3-RA3(2010) Initial characterization of the human central proteome.Burkard T.R., Planyavsky M., Kaupe I., Breitwieser F.P., Buerckstuemmer T., Bennett K.L., Superti-Furga G., Colinge J.BMC Syst. Biol. 5:17-17(2011) E3 ligase RFWD3 participates in replication checkpoint control.Gong Z., Chen J.J. Biol. Chem. 286:22308-22313(2011) RING finger and WD repeat domain 3 (RFWD3) associates with replication protein A (RPA) and facilitates RPA-mediated DNA damage response.Liu S., Chu J., Yucer N., Leng M., Wang S.Y., Chen B.P., Hittelman W.N., Wang Y.J. Biol. Chem. 286:22314-22322(2011) DNA-PK-dependent RPA2 hyperphosphorylation facilitates DNA repair and suppresses sister chromatid exchange.Liaw H., Lee D., Myung K.PLoS ONE 6:E21424-E21424(2011) Translational regulation of RPA2 via internal ribosomal entry site and by eIF3a.Yin J.Y., Dong Z.Z., Liu R.Y., Chen J., Liu Z.Q., Zhang J.T.Carcinogenesis 34:1224-1231(2013) PRP19 transforms into a sensor of RPA-ssDNA after DNA damage and drives ATR activation via a ubiquitin-mediated circuitry.Marechal A., Li J.M., Ji X.Y., Wu C.S., Yazinski S.A., Nguyen H.D., Liu S., Jimenez A.E., Jin J., Zou L.Mol. Cell 53:235-246(2014) The crystal structure of the complex of replication protein A subunits RPA32 and RPA14 reveals a mechanism for single-stranded DNA binding.Bochkarev A., Bochkareva E., Frappier L., Edwards A.M.EMBO J. 18:4498-4504(1999) Structural basis for the recognition of DNA repair proteins UNG2, XPA, and RAD52 by replication factor RPA.Mer G., Bochkarev A., Gupta R., Bochkareva E., Frappier L., Ingles C.J., Edwards A.M., Chazin W.J.Cell 103:449-456(2000) Structure of the RPA trimerization core and its role in the multistep DNA-binding mechanism of RPA.Bochkareva E., Korolev S., Lees-Miller S.P., Bochkarev A.EMBO J. 21:1855-1863(2002)

NCBI and Uniprot Product Information

• NCBI GI #: 554595586
• NCBI GeneID: 6118
• NCBI Accession #: NP_001273005.1
• NCBI GenBank Nucleotide #: NM_001286076.1
• UniProt Accession #: P15927; Q52II0; Q5TEI9; Q5TEJ5
• Molecular Weight: 55.9 kDa
• NCBI Official Full Name: replication protein A 32 kDa subunit isoform 2
• NCBI Official Synonym Full Names: replication protein A2
• NCBI Official Symbol: RPA2
• NCBI Official Synonym Symbols: REPA2; RPA32; RP-A p32; RP-A p34
• NCBI Protein Information: replication protein A 32 kDa subunit
• UniProt Protein Name: Replication protein A 32 kDa subunit
• Protein Family: Replication protein
• UniProt Gene Name: RPA2
• UniProt Synonym Gene Names: REPA2; RPA32; RPA34; RP-A p32; RF-A protein 2; RP-A p34
• UniProt Entry Name: RFA2_HUMAN

NCBI Description

This gene encodes a subunit of the heterotrimeric Replication Protein A (RPA) complex, which binds to single-stranded DNA (ssDNA), forming a nucleoprotein complex that plays an important role in DNA metabolism, being involved in DNA replication, repair, recombination, telomere maintenance, and co-ordinating the cellular response to DNA damage through activation of the ataxia telangiectasia and Rad3-related protein (ATR) kinase. The RPA complex protects single-stranded DNA from nucleases, prevents formation of secondary structures that would interfere with repair, and co-ordinates the recruitment and departure of different genome maintenance factors. The heterotrimeric complex has two different modes of ssDNA binding, a low-affinity and high-affinity mode, determined by which oligonucleotide/oligosaccharide-binding (OB) domains of the complex are utilized, and differing in the length of DNA bound. This subunit contains a single OB domain that participates in high-affinity DNA binding and also contains a winged helix domain at its carboxy terminus, which interacts with many genome maintenance protein. Post-translational modifications of the RPA complex also plays a role in co-ordinating different damage response pathways. [provided by RefSeq, Sep 2017]

Uniprot Description

RFA2: Required for DNA recombination, repair and replication. The activity of RP-A is mediated by single-stranded DNA binding and protein interactions. Required for the efficient recruitment of the DNA double-strand break repair factor RAD51 to chromatin in response to DNA damage. Heterotrimer of 70, 32 and 14 kDa chains (canonical replication protein A complex). Component of the alternative replication protein A complex (aRPA) composed of RPA1, RPA3 and RPA4. The DNA-binding activity may reside exclusively on the 70 kDa subunit. Binds to SERTAD3/RBT1. Interacts with TIPIN. Directly interacts with PPP4R2, but not with SMEK2; this interaction is DNA damage-dependent and leads RPA2 dephosphorylation by PPP4C recruitment. Interacts with RAD51, preferentially when hyperphosphorylated. Directly interacts with RFWD3. 3 isoforms of the human protein are produced by alternative splicing.

Protein type: DNA replication

Chromosomal Location of Human Ortholog: 1p35

Cellular Component: chromatin; chromosome, telomeric region; DNA replication factor A complex; nuclear chromosome, telomeric region; nucleoplasm; nucleus; PML body

Molecular Function: damaged DNA binding; enzyme binding; protein binding; protein N-terminus binding; protein phosphatase binding; single-stranded DNA binding; ubiquitin protein ligase binding

Biological Process: base-excision repair; bypass DNA synthesis; DNA damage response, detection of DNA damage; DNA repair; DNA replication; DNA strand elongation during DNA replication; double-strand break repair; double-strand break repair via homologous recombination; error-prone postreplication DNA repair; G1 DNA damage checkpoint; G1/S transition of mitotic cell cycle; mismatch repair; mitotic cell cycle; nucleotide-excision repair; nucleotide-excision repair, DNA gap filling; nucleotide-excision repair, DNA incision; telomere maintenance; telomere maintenance via recombination; telomere maintenance via semi-conservative replication; transcription-coupled nucleotide-excision repair