PRI2_MOUSE
ID PRI2_MOUSE Reviewed; 505 AA.
AC P33610; Q3U4Z3;
DT 01-FEB-1994, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1994, sequence version 1.
DT 03-AUG-2022, entry version 143.
DE RecName: Full=DNA primase large subunit;
DE AltName: Full=DNA primase 58 kDa subunit;
DE Short=p58;
GN Name=Prim2;
OS Mus musculus (Mouse).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae;
OC Murinae; Mus; Mus.
OX NCBI_TaxID=10090;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], AND PROTEIN SEQUENCE OF 30-48; 186-212 AND
RP 403-416.
RX PubMed=8463324; DOI=10.1016/s0021-9258(18)53069-6;
RA Miyazawa H., Izumi M., Tada S., Takada R., Masutani M., Ui M., Hanaoka F.;
RT "Molecular cloning of the cDNAs for the four subunits of mouse DNA
RT polymerase alpha-primase complex and their gene expression during cell
RT proliferation and the cell cycle.";
RL J. Biol. Chem. 268:8111-8122(1993).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, INTERACTION WITH PRIM1, AND
RP IDENTIFICATION IN THE DNA PRIMASE COMPLEX.
RX PubMed=8026492; DOI=10.1111/j.1432-1033.1994.tb18925.x;
RA Stadlbauer F., Brueckner A., Rehfuess C., Eckerskorn C., Lottspeich F.,
RA Foerster V., Tseng B.Y., Nasheuer H.-P.;
RT "DNA replication in vitro by recombinant DNA-polymerase-alpha-primase.";
RL Eur. J. Biochem. 222:781-793(1994).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC STRAIN=C57BL/6J, and NOD; TISSUE=Thymus;
RX PubMed=16141072; DOI=10.1126/science.1112014;
RA Carninci P., Kasukawa T., Katayama S., Gough J., Frith M.C., Maeda N.,
RA Oyama R., Ravasi T., Lenhard B., Wells C., Kodzius R., Shimokawa K.,
RA Bajic V.B., Brenner S.E., Batalov S., Forrest A.R., Zavolan M., Davis M.J.,
RA Wilming L.G., Aidinis V., Allen J.E., Ambesi-Impiombato A., Apweiler R.,
RA Aturaliya R.N., Bailey T.L., Bansal M., Baxter L., Beisel K.W., Bersano T.,
RA Bono H., Chalk A.M., Chiu K.P., Choudhary V., Christoffels A.,
RA Clutterbuck D.R., Crowe M.L., Dalla E., Dalrymple B.P., de Bono B.,
RA Della Gatta G., di Bernardo D., Down T., Engstrom P., Fagiolini M.,
RA Faulkner G., Fletcher C.F., Fukushima T., Furuno M., Futaki S.,
RA Gariboldi M., Georgii-Hemming P., Gingeras T.R., Gojobori T., Green R.E.,
RA Gustincich S., Harbers M., Hayashi Y., Hensch T.K., Hirokawa N., Hill D.,
RA Huminiecki L., Iacono M., Ikeo K., Iwama A., Ishikawa T., Jakt M.,
RA Kanapin A., Katoh M., Kawasawa Y., Kelso J., Kitamura H., Kitano H.,
RA Kollias G., Krishnan S.P., Kruger A., Kummerfeld S.K., Kurochkin I.V.,
RA Lareau L.F., Lazarevic D., Lipovich L., Liu J., Liuni S., McWilliam S.,
RA Madan Babu M., Madera M., Marchionni L., Matsuda H., Matsuzawa S., Miki H.,
RA Mignone F., Miyake S., Morris K., Mottagui-Tabar S., Mulder N., Nakano N.,
RA Nakauchi H., Ng P., Nilsson R., Nishiguchi S., Nishikawa S., Nori F.,
RA Ohara O., Okazaki Y., Orlando V., Pang K.C., Pavan W.J., Pavesi G.,
RA Pesole G., Petrovsky N., Piazza S., Reed J., Reid J.F., Ring B.Z.,
RA Ringwald M., Rost B., Ruan Y., Salzberg S.L., Sandelin A., Schneider C.,
RA Schoenbach C., Sekiguchi K., Semple C.A., Seno S., Sessa L., Sheng Y.,
RA Shibata Y., Shimada H., Shimada K., Silva D., Sinclair B., Sperling S.,
RA Stupka E., Sugiura K., Sultana R., Takenaka Y., Taki K., Tammoja K.,
RA Tan S.L., Tang S., Taylor M.S., Tegner J., Teichmann S.A., Ueda H.R.,
RA van Nimwegen E., Verardo R., Wei C.L., Yagi K., Yamanishi H.,
RA Zabarovsky E., Zhu S., Zimmer A., Hide W., Bult C., Grimmond S.M.,
RA Teasdale R.D., Liu E.T., Brusic V., Quackenbush J., Wahlestedt C.,
RA Mattick J.S., Hume D.A., Kai C., Sasaki D., Tomaru Y., Fukuda S.,
RA Kanamori-Katayama M., Suzuki M., Aoki J., Arakawa T., Iida J., Imamura K.,
RA Itoh M., Kato T., Kawaji H., Kawagashira N., Kawashima T., Kojima M.,
RA Kondo S., Konno H., Nakano K., Ninomiya N., Nishio T., Okada M., Plessy C.,
RA Shibata K., Shiraki T., Suzuki S., Tagami M., Waki K., Watahiki A.,
RA Okamura-Oho Y., Suzuki H., Kawai J., Hayashizaki Y.;
RT "The transcriptional landscape of the mammalian genome.";
RL Science 309:1559-1563(2005).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC STRAIN=FVB/N; TISSUE=Mammary tumor;
RX PubMed=15489334; DOI=10.1101/gr.2596504;
RG The MGC Project Team;
RT "The status, quality, and expansion of the NIH full-length cDNA project:
RT the Mammalian Gene Collection (MGC).";
RL Genome Res. 14:2121-2127(2004).
RN [5]
RP FUNCTION, INTERACTION WITH POLA1 AND PRIM1, AND IDENTIFICATION IN DNA
RP PRIMASE COMPLEX.
RX PubMed=8253737; DOI=10.1016/s0021-9258(19)74297-5;
RA Copeland W.C., Wang T.S.;
RT "Enzymatic characterization of the individual mammalian primase subunits
RT reveals a biphasic mechanism for initiation of DNA replication.";
RL J. Biol. Chem. 268:26179-26189(1993).
RN [6]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC TISSUE=Spleen, and Testis;
RX PubMed=21183079; DOI=10.1016/j.cell.2010.12.001;
RA Huttlin E.L., Jedrychowski M.P., Elias J.E., Goswami T., Rad R.,
RA Beausoleil S.A., Villen J., Haas W., Sowa M.E., Gygi S.P.;
RT "A tissue-specific atlas of mouse protein phosphorylation and expression.";
RL Cell 143:1174-1189(2010).
CC -!- FUNCTION: Regulatory subunit of the DNA primase complex and component
CC of the DNA polymerase alpha complex (also known as the alpha DNA
CC polymerase-primase complex) which play an essential role in the
CC initiation of DNA synthesis (PubMed:8026492, PubMed:8253737). During
CC the S phase of the cell cycle, the DNA polymerase alpha complex
CC (composed of a catalytic subunit POLA1, an accessory subunit POLA2 and
CC two primase subunits, the catalytic subunit PRIM1 and the regulatory
CC subunit PRIM2) is recruited to DNA at the replicative forks via direct
CC interactions with MCM10 and WDHD1 (By similarity). The primase subunit
CC of the polymerase alpha complex initiates DNA synthesis by
CC oligomerising short RNA primers on both leading and lagging strands
CC (PubMed:8253737). These primers are initially extended by the
CC polymerase alpha catalytic subunit and subsequently transferred to
CC polymerase delta and polymerase epsilon for processive synthesis on the
CC lagging and leading strand, respectively (By similarity). In the
CC primase complex, both subunits are necessary for the initial di-
CC nucleotide formation, but the extension of the primer depends only on
CC the catalytic subunit (PubMed:8253737). Binds RNA:DNA duplex and
CC coordinates the catalytic activities of PRIM1 and POLA2 during primase-
CC to-polymerase switch. {ECO:0000250|UniProtKB:P09884,
CC ECO:0000250|UniProtKB:P49643, ECO:0000269|PubMed:8026492,
CC ECO:0000269|PubMed:8253737}.
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000250|UniProtKB:P49643};
CC Note=Binds 1 [4Fe-4S] cluster. {ECO:0000250|UniProtKB:P49643};
CC -!- SUBUNIT: Heterodimer of a catalytic subunit PRIM1 and a regulatory
CC subunit PRIM2, also known as the DNA primase complex (PubMed:8026492,
CC PubMed:8253737). Interacts via (C-terminus) with PRIM1 (By similarity).
CC Component of the alpha DNA polymerase complex (also known as the alpha
CC DNA polymerase-primase complex) consisting of four subunits: the
CC catalytic subunit POLA1, the regulatory subunit POLA2, and the primase
CC complex subunits PRIM1 and PRIM2 respectively (PubMed:8253737). Within
CC the complex, POLA1 directly interacts with PRIM2 (PubMed:8026492).
CC {ECO:0000250|UniProtKB:P49643, ECO:0000269|PubMed:8026492,
CC ECO:0000269|PubMed:8253737}.
CC -!- DOMAIN: The RNA:DNA duplex-binding domain interacts with the template
CC phosphates at positions -2, -1, 1, and 2 positioning its bases -1, 1,
CC and 2 for duplex formation. Interacts only with the beta- and gamma-
CC phosphates of triphosphate moiety of initiating NTP of the primer. The
CC side chain of His-303 mimics a RNA base that would be paired with the
CC template nucleotide at position -1 via a hydrogen bond, thereby
CC facilitating the stacking of the initiating NTP. In the initiating
CC primosome a 'mini RNA:DNA' duplex is formed comprising three template
CC nucleotides at positions -1, 1, and 2 on one strand and His-303,
CC initiating NTP, and incoming NTP on the other strand.
CC {ECO:0000250|UniProtKB:P49643}.
CC -!- DOMAIN: The interdomain linker provides flexibility in movement
CC relative to primosome platform composed of PRIM1, the N-terminus of
CC PRIM2, the C-terminus of POLA1 and POLA2. Together with POLA1
CC interdomain linker, allows for large-scale conformational changes of
CC primosome and coordinated autoregulation of catalytic centers of PRIM1
CC and POLA1. It is proposed to move the C-terminus of PRIM2 close to
CC PRIM1 during initiation, then move it away with the 5'-end of the
CC nascent primer during elongation. The steric hindrance between the
CC N- and C-terminus of PRIM2 as the RNA primer is elongated limits its
CC length to 9 nucleotides. Ultimately a large rotation of the C-terminus
CC of PRIM2 transfers the primer to POLA1 active site for DNA synthesis.
CC {ECO:0000250|UniProtKB:P49643}.
CC -!- SIMILARITY: Belongs to the eukaryotic-type primase large subunit
CC family. {ECO:0000305}.
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DR EMBL; D13545; BAA02745.1; -; mRNA.
DR EMBL; D17385; BAA04203.1; -; mRNA.
DR EMBL; AK076095; BAC36179.1; -; mRNA.
DR EMBL; AK077760; BAC36996.1; -; mRNA.
DR EMBL; AK153970; BAE32287.1; -; mRNA.
DR EMBL; BC019500; AAH19500.1; -; mRNA.
DR CCDS; CCDS14862.1; -.
DR PIR; C46642; C46642.
DR RefSeq; NP_032948.1; NM_008922.2.
DR AlphaFoldDB; P33610; -.
DR SMR; P33610; -.
DR BioGRID; 202362; 8.
DR ComplexPortal; CPX-2088; DNA polymerase alpha:primase complex.
DR CORUM; P33610; -.
DR IntAct; P33610; 6.
DR STRING; 10090.ENSMUSP00000027312; -.
DR iPTMnet; P33610; -.
DR PhosphoSitePlus; P33610; -.
DR EPD; P33610; -.
DR MaxQB; P33610; -.
DR PaxDb; P33610; -.
DR PeptideAtlas; P33610; -.
DR PRIDE; P33610; -.
DR ProteomicsDB; 291592; -.
DR Antibodypedia; 31147; 164 antibodies from 26 providers.
DR DNASU; 19076; -.
DR Ensembl; ENSMUST00000027312; ENSMUSP00000027312; ENSMUSG00000026134.
DR GeneID; 19076; -.
DR KEGG; mmu:19076; -.
DR UCSC; uc007anq.2; mouse.
DR CTD; 5558; -.
DR MGI; MGI:97758; Prim2.
DR VEuPathDB; HostDB:ENSMUSG00000026134; -.
DR eggNOG; KOG2267; Eukaryota.
DR GeneTree; ENSGT00390000009790; -.
DR HOGENOM; CLU_026253_2_0_1; -.
DR InParanoid; P33610; -.
DR OMA; KTSMPLC; -.
DR OrthoDB; 479347at2759; -.
DR PhylomeDB; P33610; -.
DR TreeFam; TF105893; -.
DR Reactome; R-MMU-113501; Inhibition of replication initiation of damaged DNA by RB1/E2F1.
DR Reactome; R-MMU-174411; Polymerase switching on the C-strand of the telomere.
DR Reactome; R-MMU-174430; Telomere C-strand synthesis initiation.
DR Reactome; R-MMU-68952; DNA replication initiation.
DR Reactome; R-MMU-68962; Activation of the pre-replicative complex.
DR Reactome; R-MMU-69091; Polymerase switching.
DR Reactome; R-MMU-69166; Removal of the Flap Intermediate.
DR Reactome; R-MMU-69183; Processive synthesis on the lagging strand.
DR BioGRID-ORCS; 19076; 26 hits in 74 CRISPR screens.
DR ChiTaRS; Prim2; mouse.
DR PRO; PR:P33610; -.
DR Proteomes; UP000000589; Chromosome 1.
DR RNAct; P33610; protein.
DR Bgee; ENSMUSG00000026134; Expressed in fetal liver hematopoietic progenitor cell and 265 other tissues.
DR Genevisible; P33610; MM.
DR GO; GO:0005658; C:alpha DNA polymerase:primase complex; IDA:UniProtKB.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; ISS:UniProtKB.
DR GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR GO; GO:0071667; F:DNA/RNA hybrid binding; ISS:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0006270; P:DNA replication initiation; IDA:ComplexPortal.
DR GO; GO:0006269; P:DNA replication, synthesis of RNA primer; IDA:UniProtKB.
DR GO; GO:1903934; P:positive regulation of DNA primase activity; ISS:UniProtKB.
DR CDD; cd07322; PriL_PriS_Eukaryotic; 1.
DR InterPro; IPR016558; DNA_primase_lsu_euk.
DR InterPro; IPR007238; DNA_primase_lsu_euk/arc.
DR PANTHER; PTHR10537; PTHR10537; 1.
DR Pfam; PF04104; DNA_primase_lrg; 1.
DR PIRSF; PIRSF009449; DNA_primase_large_subunit; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Direct protein sequencing; DNA replication; DNA-binding; Iron;
KW Iron-sulfur; Metal-binding; Phosphoprotein; Primosome; Reference proteome.
FT CHAIN 1..505
FT /note="DNA primase large subunit"
FT /id="PRO_0000046769"
FT REGION 253..270
FT /note="Interdomain linker"
FT /evidence="ECO:0000250|UniProtKB:P49643"
FT REGION 266..503
FT /note="Interacts with PRIM1"
FT /evidence="ECO:0000250|UniProtKB:P49643"
FT REGION 300..442
FT /note="RNA:DNA duplex binding"
FT /evidence="ECO:0000250|UniProtKB:P49643"
FT REGION 463..486
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 470..486
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT BINDING 287
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:P49643"
FT BINDING 367
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:P49643"
FT BINDING 384
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:P49643"
FT BINDING 424
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:P49643"
FT MOD_RES 470
FT /note="Phosphothreonine"
FT /evidence="ECO:0000250|UniProtKB:P49643"
FT CONFLICT 109
FT /note="A -> G (in Ref. 2; BAA04203)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 505 AA; 58408 MW; 36A64C77669D8C24 CRC64;
MQFSGRIRKK LRLAGDQRNA SYPHSLQFYL QPPTENISLT EFENLAFDRV KLLKAIENLG
VSYVKGTEQY QSKLEAEIRK LKFSYRENLE DEYEPRRRDH ISHFILRLAY CQSEDLRRWF
IQQEMDLLRF RFSILPKDKV QSFLKDSHLH FEAISDEEKT LREQDIMASS PSLSGIKLES
ESVYKVPFAD ALDLFRGRKV YLEDGFAYVP LKDIVAIILN EFRATLSKAL ALTARSLPAV
QSDERLQPLL NHLSHSYTGQ DYSTQKNTGK ISLDQIDSLS TKSFPPCMRQ LHKALRENHH
LRHGGRMQYG LFLKGIGLTL EQALQFWKQE FIRGKMDPDK FDKGYSYNIR HSFGKEGKRT
DYTPFSCMKI ILTNPPGQGD YHGCPFRHSD AELLKQKMQS YKIPASGISQ ILDLVKGNHY
QVACQKYFEM THNVDDCGFS LNHPNQFFFE SQRILTGGKD IKKEISQPET PQHKPSTQKT
RDAASALASL DSSLEMDLEG LEEYF
