PRI2_YEAST
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 03-AUG-2022, entry version 173.
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 PDBsum; 3LGB; -.
DR PDBsum; 6DTV; -.
DR PDBsum; 6DTZ; -.
DR PDBsum; 6DU0; -.
DR AlphaFoldDB; P20457; -.
DR SMR; P20457; -.
DR BioGRID; 34088; 235.
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; P20457; -.
DR PRIDE; P20457; -.
DR EnsemblFungi; YKL045W_mRNA; YKL045W; YKL045W.
DR GeneID; 853821; -.
DR KEGG; sce:YKL045W; -.
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; KTSMPLC; -.
DR BioCyc; YEAST:G3O-31846-MON; -.
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 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; IC:SGD.
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 GO; GO:0006273; P:lagging strand elongation; IC:SGD.
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 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 S-
FT 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 S-
FT 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
