PRI1_MOUSE
ID PRI1_MOUSE Reviewed; 417 AA.
AC P20664;
DT 01-FEB-1991, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1991, sequence version 1.
DT 03-AUG-2022, entry version 154.
DE RecName: Full=DNA primase small subunit;
DE EC=2.7.7.102 {ECO:0000269|PubMed:8026492, ECO:0000269|PubMed:8253737};
DE AltName: Full=DNA primase 49 kDa subunit;
DE Short=p49;
GN Name=Prim1;
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 PARTIAL PROTEIN SEQUENCE.
RC TISSUE=Lymphoid tissue;
RX PubMed=2925677; DOI=10.1016/s0021-9258(18)83684-5;
RA Prussak C.E., Almazan M.T., Tseng B.Y.;
RT "Mouse primase p49 subunit molecular cloning indicates conserved and
RT divergent regions.";
RL J. Biol. Chem. 264:4957-4963(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], AND PROTEIN SEQUENCE OF 51-66 AND 222-237.
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 [3]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, INTERACTION WITH PRIM2, AND IDENTIFICATION IN DNA PRIMASE
RP 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 [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND INTERACTION WITH PRIM2.
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 [5]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC TISSUE=Spleen;
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: Catalytic subunit of the DNA primase complex and component of
CC 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:8253737, PubMed:8026492). 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 These primers are initially extended by the polymerase alpha catalytic
CC subunit and subsequently transferred to polymerase delta and polymerase
CC epsilon for processive synthesis on the lagging and leading strand,
CC respectively (By similarity). In the primase complex, both subunits are
CC necessary for the initial di-nucleotide formation, but the extension of
CC the primer depends only on the catalytic subunit (PubMed:8253737). Can
CC add both ribo- and deoxynucleotides during elongation of the primers
CC (PubMed:8253737). Synthesizes 9-mer RNA primers (also known as the
CC 'unit length' RNA primers) (By similarity). Incorporates only
CC ribonucleotides in the presence of ribo- and deoxy-nucleotide
CC triphosphates (rNTPs, dNTPs). Requires template thymine or cytidine to
CC start the RNA primer synthesis, with an adenine or guanine at its 5'-
CC end (By similarity). Binds single stranded DNA (PubMed:8253737).
CC {ECO:0000250|UniProtKB:P09884, ECO:0000250|UniProtKB:P49642,
CC ECO:0000269|PubMed:8026492, ECO:0000269|PubMed:8253737}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ssDNA + n NTP = ssDNA/pppN(pN)n-1 hybrid + (n-1) diphosphate.;
CC EC=2.7.7.102; Evidence={ECO:0000269|PubMed:8026492,
CC ECO:0000269|PubMed:8253737};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:P49642};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000250|UniProtKB:P49642};
CC -!- ACTIVITY REGULATION: The presence of the regulatory subunit PRIM2/p58
CC accelerates the kinetics of initiation and primer extension.
CC {ECO:0000269|PubMed:8253737}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.25 mM for ATP {ECO:0000269|PubMed:8253737};
CC KM=3 mM for ATP (in presence of the regulatory PRIM2/p58)
CC {ECO:0000269|PubMed:8253737};
CC KM=4.1 uM for oligo(A)-primed poly(dT) {ECO:0000269|PubMed:8253737};
CC -!- SUBUNIT: Heterodimer of a catalytic subunit PRIM1 and a regulatory
CC subunit PRIM2, also known as the DNA primase complex (PubMed:8253737,
CC PubMed:8026492). Interacts with PRIM2/p58 (via C-terminus) (By
CC similarity). Component of the alpha DNA polymerase complex (also known
CC as the alpha DNA polymerase-primase complex) consisting of four
CC subunits: the catalytic subunit POLA1, the regulatory subunit POLA2,
CC and the primase complex subunits PRIM1 and PRIM2 respectively
CC (PubMed:8253737). Within the complex, POLA1 directly interacts with
CC PRIM2 (PubMed:8253737). {ECO:0000250|UniProtKB:P49642,
CC ECO:0000269|PubMed:8026492, ECO:0000269|PubMed:8253737}.
CC -!- INTERACTION:
CC P20664; P33609: Pola1; NbExp=4; IntAct=EBI-848742, EBI-688051;
CC -!- DOMAIN: The catalytic domain (residues 1-190 and 303-408) adopts a
CC typical 'prim' fold structure formed by two three strand beta-sheets
CC that line the inside of the lower and upper parts, each surrounded by
CC alpha-helices on the outside. It comprises a highly conserved catalytic
CC triad, a structural zinc-binding motif and the nucleotide-binding
CC motifs. The Asp-109, Asp-111 and Asp-305 catalytic triad binds two Mn2+
CC or Mg2+ ions which activate for nucleophilic attack the 3'-hydroxyl of
CC the growing RNA primer or of the first NTP bound at the initiation
CC site. The nucleotide-binding motifs coordinate the phosphates, the
CC ribose and the base of a NTP molecule. The interaction between O2' of
CC the initiating NTP and Asp-305 stabilizes the ribose during the di-
CC nucleotide synthesis. It is proposed that the first nucleotide binds to
CC the elongation site, followed by binding to the initiation site of a
CC second NTP, which will become the 5'-terminal nucleotide of the primer.
CC {ECO:0000250|UniProtKB:P49642}.
CC -!- MISCELLANEOUS: The bound zinc ion is not a cofactor. It is bound to a
CC zinc knuckle motif that may be involved in sequence recognition and the
CC binding of ssDNA (By similarity). {ECO:0000250}.
CC -!- SIMILARITY: Belongs to the eukaryotic-type primase small subunit
CC family. {ECO:0000305}.
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DR EMBL; J04620; AAA39880.1; -; mRNA.
DR EMBL; D13544; BAA02744.1; -; mRNA.
DR CCDS; CCDS36086.1; -.
DR PIR; A33269; A33269.
DR RefSeq; NP_032947.1; NM_008921.2.
DR AlphaFoldDB; P20664; -.
DR SMR; P20664; -.
DR BioGRID; 202361; 7.
DR ComplexPortal; CPX-2088; DNA polymerase alpha:primase complex.
DR CORUM; P20664; -.
DR IntAct; P20664; 4.
DR STRING; 10090.ENSMUSP00000026461; -.
DR iPTMnet; P20664; -.
DR PhosphoSitePlus; P20664; -.
DR EPD; P20664; -.
DR PaxDb; P20664; -.
DR PeptideAtlas; P20664; -.
DR PRIDE; P20664; -.
DR ProteomicsDB; 291558; -.
DR DNASU; 19075; -.
DR Ensembl; ENSMUST00000026461; ENSMUSP00000026461; ENSMUSG00000025395.
DR GeneID; 19075; -.
DR KEGG; mmu:19075; -.
DR UCSC; uc011xpx.2; mouse.
DR CTD; 5557; -.
DR MGI; MGI:97757; Prim1.
DR VEuPathDB; HostDB:ENSMUSG00000025395; -.
DR eggNOG; KOG2851; Eukaryota.
DR GeneTree; ENSGT00390000011466; -.
DR InParanoid; P20664; -.
DR OMA; NVTRGFN; -.
DR OrthoDB; 1346165at2759; -.
DR PhylomeDB; P20664; -.
DR TreeFam; TF312823; -.
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; 19075; 23 hits in 75 CRISPR screens.
DR ChiTaRS; Prim1; mouse.
DR PRO; PR:P20664; -.
DR Proteomes; UP000000589; Chromosome 10.
DR RNAct; P20664; protein.
DR Bgee; ENSMUSG00000025395; Expressed in floor plate of midbrain and 247 other tissues.
DR ExpressionAtlas; P20664; baseline and differential.
DR Genevisible; P20664; MM.
DR GO; GO:0005658; C:alpha DNA polymerase:primase complex; IDA:UniProtKB.
DR GO; GO:0003896; F:DNA primase activity; IDA:UniProtKB.
DR GO; GO:0000287; F:magnesium ion binding; ISS:UniProtKB.
DR GO; GO:0032553; F:ribonucleotide binding; ISS:UniProtKB.
DR GO; GO:0008270; F:zinc ion binding; ISS:UniProtKB.
DR GO; GO:0006270; P:DNA replication initiation; IDA:ComplexPortal.
DR GO; GO:0006269; P:DNA replication, synthesis of RNA primer; IDA:UniProtKB.
DR CDD; cd04860; AE_Prim_S; 1.
DR InterPro; IPR002755; DNA_primase_S.
DR InterPro; IPR014052; DNA_primase_ssu_euk/arc.
DR PANTHER; PTHR10536; PTHR10536; 1.
DR Pfam; PF01896; DNA_primase_S; 1.
DR TIGRFAMs; TIGR00335; primase_sml; 1.
PE 1: Evidence at protein level;
KW Acetylation; Direct protein sequencing; DNA replication;
KW DNA-directed RNA polymerase; Magnesium; Manganese; Metal-binding;
KW Nucleotidyltransferase; Primosome; Reference proteome; Transcription;
KW Transferase; Zinc.
FT CHAIN 1..417
FT /note="DNA primase small subunit"
FT /id="PRO_0000046731"
FT MOTIF 121..131
FT /note="Zinc knuckle motif"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT ACT_SITE 44
FT /evidence="ECO:0000255"
FT ACT_SITE 109
FT /evidence="ECO:0000255"
FT ACT_SITE 111
FT /evidence="ECO:0000255"
FT BINDING 109..111
FT /ligand="a ribonucleoside 5'-triphosphate"
FT /ligand_id="ChEBI:CHEBI:61557"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 109
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 109
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 109
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 109
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 111
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 111
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 111
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 111
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 121
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 122
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 128
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 131
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 160..166
FT /ligand="a ribonucleoside 5'-triphosphate"
FT /ligand_id="ChEBI:CHEBI:61557"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 305
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 305
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 314..317
FT /ligand="a ribonucleoside 5'-triphosphate"
FT /ligand_id="ChEBI:CHEBI:61557"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT BINDING 323
FT /ligand="a ribonucleoside 5'-triphosphate"
FT /ligand_id="ChEBI:CHEBI:61557"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT MOD_RES 1
FT /note="N-acetylmethionine"
FT /evidence="ECO:0000250|UniProtKB:P49642"
FT CONFLICT 57
FT /note="Y -> I (in Ref. 1; AAA39880)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 417 AA; 49295 MW; 54EBF4DA4DE47D8A CRC64;
MEPFDPAELP ELLKLYYRRL FPYAQYYRWL NYGGVTKNYF QHREFSFTLK DDIYIRYQSF
NNQSELEKEM QKMNPYKIDI GAVYSHRPNQ HNTVKLGAFQ AQEKELVFDI DMTDYDDVRR
CCSSADICSK CWTLMTMAMR IIDRALKEDF GFKHRLWVYS GRRGVHCWVC DESVRKLSSA
VRSGIVEYLS LVKGGQDVKK KVHLNEKVHP FVRKSINIIK KYFEEYALVG QDILENKENW
DKILALVPET IHDELQRGFQ KFHSSPQRWE HLRKVANSSQ NMKNDKCGPW LEWEVMLQYC
FPRLDVNVSK GVNHLLKSPF SVHPKTGRIS VPIDFHKVDQ FDPFTVPTIS AICRELDMVS
THEKEKEENE ADSKHRVRGY KKTSLAPYVK VFEQFLENLD KSRKGELLKK SDLQKDF
