ID   PRI2_YEAST              Reviewed;         528 AA.
AC   P20457; D6VXP2;
DT   01-FEB-1991, integrated into UniProtKB/Swiss-Prot.
DT   01-FEB-1991, sequence version 1.
DT   27-MAR-2024, entry version 181.
DE   RecName: Full=DNA primase large subunit;
DE   AltName: Full=DNA polymerase alpha:primase complex p58 subunit;
DE            Short=DNA polymerase-primase complex p58 subunit;
DE            Short=Pol alpha-primase complex p58 subunit;
DE   AltName: Full=DNA primase 58 kDa subunit;
GN   Name=PRI2; OrderedLocusNames=YKL045W; ORFNames=YKL258;
OS   Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast).
OC   Eukaryota; Fungi; Dikarya; Ascomycota; Saccharomycotina; Saccharomycetes;
OC   Saccharomycetales; Saccharomycetaceae; Saccharomyces.
OX   NCBI_TaxID=559292;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX   PubMed=2528682; DOI=10.1128/mcb.9.7.3081-3087.1989;
RA   Foiani M., Santocanale C., Plevani P., Lucchini G.;
RT   "A single essential gene, PRI2, encodes the large subunit of DNA primase in
RT   Saccharomyces cerevisiae.";
RL   Mol. Cell. Biol. 9:3081-3087(1989).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=ATCC 204508 / S288c;
RX   PubMed=8154189; DOI=10.1002/yea.320091212;
RA   Purnelle B., Tettelin H., van Dyck L., Skala J., Goffeau A.;
RT   "The sequence of a 17.5 kb DNA fragment on the left arm of yeast chromosome
RT   XI identifies the protein kinase gene ELM1, the DNA primase gene PRI2, a
RT   new gene encoding a putative histone and seven new open reading frames.";
RL   Yeast 9:1379-1384(1993).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=ATCC 204508 / S288c;
RX   PubMed=8196765; DOI=10.1038/369371a0;
RA   Dujon B., Alexandraki D., Andre B., Ansorge W., Baladron V.,
RA   Ballesta J.P.G., Banrevi A., Bolle P.-A., Bolotin-Fukuhara M., Bossier P.,
RA   Bou G., Boyer J., Buitrago M.J., Cheret G., Colleaux L.,
RA   Daignan-Fornier B., del Rey F., Dion C., Domdey H., Duesterhoeft A.,
RA   Duesterhus S., Entian K.-D., Erfle H., Esteban P.F., Feldmann H.,
RA   Fernandes L., Fobo G.M., Fritz C., Fukuhara H., Gabel C., Gaillon L.,
RA   Garcia-Cantalejo J.M., Garcia-Ramirez J.J., Gent M.E., Ghazvini M.,
RA   Goffeau A., Gonzalez A., Grothues D., Guerreiro P., Hegemann J.H.,
RA   Hewitt N., Hilger F., Hollenberg C.P., Horaitis O., Indge K.J.,
RA   Jacquier A., James C.M., Jauniaux J.-C., Jimenez A., Keuchel H.,
RA   Kirchrath L., Kleine K., Koetter P., Legrain P., Liebl S., Louis E.J.,
RA   Maia e Silva A., Marck C., Monnier A.-L., Moestl D., Mueller S.,
RA   Obermaier B., Oliver S.G., Pallier C., Pascolo S., Pfeiffer F.,
RA   Philippsen P., Planta R.J., Pohl F.M., Pohl T.M., Poehlmann R.,
RA   Portetelle D., Purnelle B., Puzos V., Ramezani Rad M., Rasmussen S.W.,
RA   Remacha M.A., Revuelta J.L., Richard G.-F., Rieger M.,
RA   Rodrigues-Pousada C., Rose M., Rupp T., Santos M.A., Schwager C.,
RA   Sensen C., Skala J., Soares H., Sor F., Stegemann J., Tettelin H.,
RA   Thierry A., Tzermia M., Urrestarazu L.A., van Dyck L.,
RA   van Vliet-Reedijk J.C., Valens M., Vandenbol M., Vilela C., Vissers S.,
RA   von Wettstein D., Voss H., Wiemann S., Xu G., Zimmermann J., Haasemann M.,
RA   Becker I., Mewes H.-W.;
RT   "Complete DNA sequence of yeast chromosome XI.";
RL   Nature 369:371-378(1994).
RN   [4]
RP   GENOME REANNOTATION.
RC   STRAIN=ATCC 204508 / S288c;
RX   PubMed=24374639; DOI=10.1534/g3.113.008995;
RA   Engel S.R., Dietrich F.S., Fisk D.G., Binkley G., Balakrishnan R.,
RA   Costanzo M.C., Dwight S.S., Hitz B.C., Karra K., Nash R.S., Weng S.,
RA   Wong E.D., Lloyd P., Skrzypek M.S., Miyasato S.R., Simison M., Cherry J.M.;
RT   "The reference genome sequence of Saccharomyces cerevisiae: Then and now.";
RL   G3 (Bethesda) 4:389-398(2014).
RN   [5]
RP   COMPOSITION OF THE DNA POLYMERASE ALPHA:PRIMASE COMPLEX.
RX   PubMed=3061469; DOI=10.1016/0167-4781(88)90096-6;
RA   Plevani P., Foiani M., Muzi Falconi M., Pizzagalli A., Santocanale C.,
RA   Francesconi S., Valsasnini P., Comedini A., Piatti S., Lucchini G.;
RT   "The yeast DNA polymerase-primase complex: genes and proteins.";
RL   Biochim. Biophys. Acta 951:268-273(1988).
RN   [6]
RP   MUTAGENESIS.
RX   PubMed=2023935; DOI=10.1073/pnas.88.9.3877;
RA   Francesconi S., Longhese M.P., Piseri A., Santocanale C., Lucchini G.,
RA   Plevani P.;
RT   "Mutations in conserved yeast DNA primase domains impair DNA replication in
RT   vivo.";
RL   Proc. Natl. Acad. Sci. U.S.A. 88:3877-3881(1991).
RN   [7]
RP   INTERACTION WITH MCM10.
RX   PubMed=16675460; DOI=10.1074/jbc.m513551200;
RA   Ricke R.M., Bielinsky A.-K.;
RT   "A conserved Hsp10-like domain in Mcm10 is required to stabilize the
RT   catalytic subunit of DNA polymerase-alpha in budding yeast.";
RL   J. Biol. Chem. 281:18414-18425(2006).
RN   [8]
RP   IRON-SULFUR-BINDING, COFACTOR, AND MUTAGENESIS OF CYS-336; CYS-417; CYS-434
RP   AND CYS-474.
RX   PubMed=17704817; DOI=10.1038/nsmb1288;
RA   Klinge S., Hirst J., Maman J.D., Krude T., Pellegrini L.;
RT   "An iron-sulfur domain of the eukaryotic primase is essential for RNA
RT   primer synthesis.";
RL   Nat. Struct. Mol. Biol. 14:875-877(2007).
RN   [9]
RP   X-RAY CRYSTALLOGRAPHY (1.54 ANGSTROMS) OF 317-512 IN COMPLEX WITH
RP   IRON-SULFUR.
RX   PubMed=20404922; DOI=10.1371/journal.pone.0010083;
RA   Sauguet L., Klinge S., Perera R.L., Maman J.D., Pellegrini L.;
RT   "Shared active site architecture between the large subunit of eukaryotic
RT   primase and DNA photolyase.";
RL   PLoS ONE 5:E10083-E10083(2010).
CC   -!- FUNCTION: DNA primase is the polymerase that synthesizes small RNA
CC       primers for the Okazaki fragments made during discontinuous DNA
CC       replication. In a complex with DNA polymerase alpha (DNA polymerase
CC       alpha:primase) constitutes a replicative polymerase. Both primase
CC       components participate in formation of the active center, but the ATP-
CC       binding site is exclusively located on p48.
CC   -!- COFACTOR:
CC       Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC         Evidence={ECO:0000269|PubMed:17704817};
CC       Note=Binds 1 [4Fe-4S] cluster. {ECO:0000269|PubMed:17704817};
CC   -!- SUBUNIT: DNA polymerase alpha:primase is a four subunit enzyme complex,
CC       which is assembled throughout the cell cycle, and consists of the two
CC       DNA polymerase subunits A POL1 and B POL12, and the DNA primase large
CC       PRI2 and small PRI1 subunits (PubMed:3061469). Interacts with MCM10
CC       (PubMed:16675460). {ECO:0000269|PubMed:16675460,
CC       ECO:0000269|PubMed:3061469}.
CC   -!- DEVELOPMENTAL STAGE: Fluctuates in amount during the cell cycle.
CC   -!- SIMILARITY: Belongs to the eukaryotic-type primase large subunit
CC       family. {ECO:0000305}.
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DR   EMBL; M27209; AAA34900.1; -; Genomic_DNA.
DR   EMBL; X71621; CAA50626.1; -; Genomic_DNA.
DR   EMBL; Z28045; CAA81880.1; -; Genomic_DNA.
DR   EMBL; BK006944; DAA09112.1; -; Genomic_DNA.
DR   PIR; A32497; A32497.
DR   RefSeq; NP_012879.1; NM_001179611.1.
DR   PDB; 3LGB; X-ray; 1.54 A; A/B=317-512.
DR   PDB; 6DTV; X-ray; 1.12 A; A=316-512.
DR   PDB; 6DTZ; X-ray; 1.36 A; A=316-512.
DR   PDB; 6DU0; X-ray; 1.82 A; A=316-512.
DR   PDB; 7TL2; X-ray; 1.53 A; A=316-512.
DR   PDB; 7TL3; X-ray; 2.07 A; A=316-512.
DR   PDB; 7TL4; X-ray; 1.80 A; A=316-512.
DR   PDB; 8B9A; EM; 3.50 A; A=1-528.
DR   PDB; 8B9B; EM; 3.50 A; A=1-528.
DR   PDB; 8B9C; EM; 4.60 A; A=1-528.
DR   PDBsum; 3LGB; -.
DR   PDBsum; 6DTV; -.
DR   PDBsum; 6DTZ; -.
DR   PDBsum; 6DU0; -.
DR   PDBsum; 7TL2; -.
DR   PDBsum; 7TL3; -.
DR   PDBsum; 7TL4; -.
DR   PDBsum; 8B9A; -.
DR   PDBsum; 8B9B; -.
DR   PDBsum; 8B9C; -.
DR   AlphaFoldDB; P20457; -.
DR   EMDB; EMD-15924; -.
DR   SMR; P20457; -.
DR   BioGRID; 34088; 239.
DR   ComplexPortal; CPX-2091; DNA polymerase alpha:primase complex.
DR   DIP; DIP-2535N; -.
DR   IntAct; P20457; 15.
DR   MINT; P20457; -.
DR   STRING; 4932.YKL045W; -.
DR   MaxQB; P20457; -.
DR   PaxDb; 4932-YKL045W; -.
DR   PeptideAtlas; P20457; -.
DR   EnsemblFungi; YKL045W_mRNA; YKL045W; YKL045W.
DR   GeneID; 853821; -.
DR   KEGG; sce:YKL045W; -.
DR   AGR; SGD:S000001528; -.
DR   SGD; S000001528; PRI2.
DR   VEuPathDB; FungiDB:YKL045W; -.
DR   eggNOG; KOG2267; Eukaryota.
DR   GeneTree; ENSGT00390000009790; -.
DR   HOGENOM; CLU_026253_1_0_1; -.
DR   InParanoid; P20457; -.
DR   OMA; RINYKPW; -.
DR   OrthoDB; 3184472at2759; -.
DR   BioCyc; YEAST:G3O-31846-MONOMER; -.
DR   Reactome; R-SCE-113501; Inhibition of replication initiation of damaged DNA by RB1/E2F1.
DR   Reactome; R-SCE-68952; DNA replication initiation.
DR   Reactome; R-SCE-68962; Activation of the pre-replicative complex.
DR   Reactome; R-SCE-69091; Polymerase switching.
DR   Reactome; R-SCE-69166; Removal of the Flap Intermediate.
DR   Reactome; R-SCE-69183; Processive synthesis on the lagging strand.
DR   BioGRID-ORCS; 853821; 0 hits in 10 CRISPR screens.
DR   EvolutionaryTrace; P20457; -.
DR   PRO; PR:P20457; -.
DR   Proteomes; UP000002311; Chromosome XI.
DR   RNAct; P20457; Protein.
DR   GO; GO:0005658; C:alpha DNA polymerase:primase complex; IDA:SGD.
DR   GO; GO:0005635; C:nuclear envelope; IDA:SGD.
DR   GO; GO:0005634; C:nucleus; IDA:SGD.
DR   GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IEA:UniProtKB-KW.
DR   GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR   GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR   GO; GO:0006260; P:DNA replication; IMP:SGD.
DR   GO; GO:0006270; P:DNA replication initiation; IBA:GO_Central.
DR   GO; GO:0006269; P:DNA replication, synthesis of RNA primer; IDA:SGD.
DR   GO; GO:0006302; P:double-strand break repair; IMP:SGD.
DR   CDD; cd07322; PriL_PriS_Eukaryotic; 1.
DR   Gene3D; 1.20.930.80; -; 1.
DR   InterPro; IPR016558; DNA_primase_lsu_euk.
DR   InterPro; IPR007238; DNA_primase_lsu_euk/arc.
DR   PANTHER; PTHR10537; DNA PRIMASE LARGE SUBUNIT; 1.
DR   PANTHER; PTHR10537:SF3; DNA PRIMASE LARGE SUBUNIT; 1.
DR   Pfam; PF04104; DNA_primase_lrg; 1.
DR   PIRSF; PIRSF009449; DNA_primase_large_subunit; 1.
PE   1: Evidence at protein level;
KW   3D-structure; 4Fe-4S; DNA replication; DNA-binding; Iron; Iron-sulfur;
KW   Metal-binding; Primosome; Reference proteome.
FT   CHAIN           1..528
FT                   /note="DNA primase large subunit"
FT                   /id="PRO_0000046776"
FT   REGION          210..239
FT                   /note="H-T-H-like motif"
FT                   /evidence="ECO:0000255"
FT   BINDING         336
FT                   /ligand="[4Fe-4S] cluster"
FT                   /ligand_id="ChEBI:CHEBI:49883"
FT                   /evidence="ECO:0000269|PubMed:20404922"
FT   BINDING         417
FT                   /ligand="[4Fe-4S] cluster"
FT                   /ligand_id="ChEBI:CHEBI:49883"
FT                   /evidence="ECO:0000269|PubMed:20404922"
FT   BINDING         434
FT                   /ligand="[4Fe-4S] cluster"
FT                   /ligand_id="ChEBI:CHEBI:49883"
FT                   /evidence="ECO:0000269|PubMed:20404922"
FT   BINDING         474
FT                   /ligand="[4Fe-4S] cluster"
FT                   /ligand_id="ChEBI:CHEBI:49883"
FT                   /evidence="ECO:0000269|PubMed:20404922"
FT   MUTAGEN         152
FT                   /note="E->Q: Temperature-sensitive."
FT                   /evidence="ECO:0000269|PubMed:2023935"
FT   MUTAGEN         336
FT                   /note="C->S: Mild disruption of iron-sulfur-binding. Strong
FT                   disruption of iron-sulfur-binding; when associated with
FT                   S-474. Strong disruption of iron-sulfur-binding, leading to
FT                   destabilization of the protein and preventing its
FT                   purification; when associated with S-417 or S-434."
FT                   /evidence="ECO:0000269|PubMed:17704817"
FT   MUTAGEN         401
FT                   /note="H->S: Lethal."
FT                   /evidence="ECO:0000269|PubMed:2023935"
FT   MUTAGEN         417
FT                   /note="C->S: Mild disruption of iron-sulfur-binding. Strong
FT                   disruption of iron-sulfur-binding, leading to
FT                   destabilization of the protein and preventing its
FT                   purification; when associated with S-336."
FT                   /evidence="ECO:0000269|PubMed:17704817"
FT   MUTAGEN         434
FT                   /note="C->S: Mild disruption of iron-sulfur-binding. Strong
FT                   disruption of iron-sulfur-binding, leading to
FT                   destabilization of the protein and preventing its
FT                   purification; when associated with S-336."
FT                   /evidence="ECO:0000269|PubMed:17704817"
FT   MUTAGEN         434
FT                   /note="C->Y: Temperature-sensitive."
FT                   /evidence="ECO:0000269|PubMed:17704817"
FT   MUTAGEN         474
FT                   /note="C->S: Mild disruption of iron-sulfur-binding. Strong
FT                   disruption of iron-sulfur-binding; when associated with
FT                   S-336."
FT                   /evidence="ECO:0000269|PubMed:17704817"
FT   STRAND          318..320
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           323..325
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           327..330
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           335..347
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           352..365
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           369..381
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           382..384
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           388..394
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           396..402
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   TURN            403..406
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           417..422
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           435..438
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           441..450
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           455..466
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           470..481
FT                   /evidence="ECO:0007829|PDB:6DTV"
FT   HELIX           500..509
FT                   /evidence="ECO:0007829|PDB:6DTV"
SQ   SEQUENCE   528 AA;  62263 MW;  2DA521AA104D3D16 CRC64;
     MFRQSKRRIA SRKNFSSYDD IVKSELDVGN TNAANQIILS SSSSEEEKKL YARLYESKLS
     FYDLPPQGEI TLEQFEIWAI DRLKILLEIE SCLSRNKSIK EIETIIKPQF QKLLPFNTES
     LEDRKKDYYS HFILRLCFCR SKELREKFVR AETFLFKIRF NMLTSTDQTK FVQSLDLPLL
     QFISNEEKAE LSHQLYQTVS ASLQFQLNLN EEHQRKQYFQ QEKFIKLPFE NVIELVGNRL
     VFLKDGYAYL PQFQQLNLLS NEFASKLNQE LIKTYQYLPR LNEDDRLLPI LNHLSSGYTI
     ADFNQQKANQ FSENVDDEIN AQSVWSEEIS SNYPLCIKNL MEGLKKNHHL RYYGRQQLSL
     FLKGIGLSAD EALKFWSEAF TRNGNMTMEK FNKEYRYSFR HNYGLEGNRI NYKPWDCHTI
     LSKPRPGRGD YHGCPFRDWS HERLSAELRS MKLTQAQIIS VLDSCQKGEY TIACTKVFEM
     THNSASADLE IGEQTHIAHP NLYFERSRQL QKKQQKLEKE KLFNNGNH
//