DNAG_SACS2
ID DNAG_SACS2 Reviewed; 406 AA.
AC P95980;
DT 30-MAY-2000, integrated into UniProtKB/Swiss-Prot.
DT 01-JUL-1997, sequence version 2.
DT 03-AUG-2022, entry version 122.
DE RecName: Full=DNA primase DnaG {ECO:0000255|HAMAP-Rule:MF_00007};
DE EC=2.7.7.101 {ECO:0000255|HAMAP-Rule:MF_00007, ECO:0000269|PubMed:20122937};
GN Name=dnaG {ECO:0000255|HAMAP-Rule:MF_00007}; OrderedLocusNames=SSO0079;
GN ORFNames=C04_042, C05_030;
OS Saccharolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
OS (Sulfolobus solfataricus).
OC Archaea; Crenarchaeota; Thermoprotei; Sulfolobales; Sulfolobaceae;
OC Saccharolobus.
OX NCBI_TaxID=273057;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 35092 / DSM 1617 / JCM 11322 / P2;
RX PubMed=8899719; DOI=10.1111/j.1365-2958.1996.tb02666.x;
RA Sensen C.W., Klenk H.-P., Singh R.K., Allard G., Chan C.C.-Y., Liu Q.Y.,
RA Penny S.L., Young F., Schenk M.E., Gaasterland T., Doolittle W.F.,
RA Ragan M.A., Charlebois R.L.;
RT "Organizational characteristics and information content of an archaeal
RT genome: 156 kb of sequence from Sulfolobus solfataricus P2.";
RL Mol. Microbiol. 22:175-191(1996).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 35092 / DSM 1617 / JCM 11322 / P2;
RX PubMed=11427726; DOI=10.1073/pnas.141222098;
RA She Q., Singh R.K., Confalonieri F., Zivanovic Y., Allard G., Awayez M.J.,
RA Chan-Weiher C.C.-Y., Clausen I.G., Curtis B.A., De Moors A., Erauso G.,
RA Fletcher C., Gordon P.M.K., Heikamp-de Jong I., Jeffries A.C., Kozera C.J.,
RA Medina N., Peng X., Thi-Ngoc H.P., Redder P., Schenk M.E., Theriault C.,
RA Tolstrup N., Charlebois R.L., Doolittle W.F., Duguet M., Gaasterland T.,
RA Garrett R.A., Ragan M.A., Sensen C.W., Van der Oost J.;
RT "The complete genome of the crenarchaeon Sulfolobus solfataricus P2.";
RL Proc. Natl. Acad. Sci. U.S.A. 98:7835-7840(2001).
RN [3]
RP INTERACTION WITH EXOSOME.
RC STRAIN=ATCC 35092 / DSM 1617 / JCM 11322 / P2;
RX PubMed=12947419; DOI=10.1038/sj.embor.embor929;
RA Evguenieva-Hackenberg E., Walter P., Hochleitner E., Lottspeich F.,
RA Klug G.;
RT "An exosome-like complex in Sulfolobus solfataricus.";
RL EMBO Rep. 4:889-893(2003).
RN [4]
RP INTERACTION WITH EXOSOME.
RX PubMed=17078816; DOI=10.1111/j.1365-2958.2006.05393.x;
RA Walter P., Klein F., Lorentzen E., Ilchmann A., Klug G.,
RA Evguenieva-Hackenberg E.;
RT "Characterization of native and reconstituted exosome complexes from the
RT hyperthermophilic archaeon Sulfolobus solfataricus.";
RL Mol. Microbiol. 62:1076-1089(2006).
RN [5]
RP FUNCTION AS A PRIMASE, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBUNIT, AND MUTAGENESIS OF GLU-175.
RX PubMed=20122937; DOI=10.1016/j.jmb.2010.01.057;
RA Zuo Z., Rodgers C.J., Mikheikin A.L., Trakselis M.A.;
RT "Characterization of a functional DnaG-type primase in archaea:
RT implications for a dual-primase system.";
RL J. Mol. Biol. 397:664-676(2010).
RN [6]
RP INTERACTION WITH EXOSOME.
RC STRAIN=ATCC 35092 / DSM 1617 / JCM 11322 / P2;
RX PubMed=22503705; DOI=10.1016/j.biochi.2012.03.026;
RA Witharana C., Roppelt V., Lochnit G., Klug G., Evguenieva-Hackenberg E.;
RT "Heterogeneous complexes of the RNA exosome in Sulfolobus solfataricus.";
RL Biochimie 94:1578-1587(2012).
RN [7]
RP FUNCTION, INTERACTION WITH MCM AND DNA, AND MUTAGENESIS OF ASP-179; ASP-220
RP AND ASP-222.
RX PubMed=23357171; DOI=10.1016/j.jmb.2013.01.025;
RA Bauer R.J., Graham B.W., Trakselis M.A.;
RT "Novel interaction of the bacterial-Like DnaG primase with the MCM helicase
RT in archaea.";
RL J. Mol. Biol. 425:1259-1273(2013).
RN [8]
RP FUNCTION, AND INTERACTION WITH CSL4.
RX PubMed=23324612; DOI=10.4161/rna.23450;
RA Hou L., Klug G., Evguenieva-Hackenberg E.;
RT "The archaeal DnaG protein needs Csl4 for binding to the exosome and
RT enhances its interaction with adenine-rich RNAs.";
RL RNA Biol. 10:415-424(2013).
CC -!- FUNCTION: RNA polymerase that catalyzes the synthesis of short RNA
CC molecules used as primers for DNA polymerase during DNA replication.
CC Can use NTPs but not dNTPs. Binds DNA. Also part of the exosome, which
CC is a complex involved in RNA degradation. Acts as a poly(A)-binding
CC protein that enhances the interaction between heteromeric, adenine-rich
CC transcripts and the exosome. {ECO:0000255|HAMAP-Rule:MF_00007,
CC ECO:0000269|PubMed:20122937, ECO:0000269|PubMed:23324612,
CC ECO:0000269|PubMed:23357171}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ssDNA + n NTP = ssDNA/pppN(pN)n-1 hybrid + (n-1) diphosphate.;
CC EC=2.7.7.101; Evidence={ECO:0000255|HAMAP-Rule:MF_00007,
CC ECO:0000269|PubMed:20122937};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00007};
CC Note=Binds two Mg(2+) per subunit. {ECO:0000255|HAMAP-Rule:MF_00007};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=85 uM for NTPs {ECO:0000269|PubMed:20122937};
CC Vmax=0.72 pmol/min/ug enzyme {ECO:0000269|PubMed:20122937};
CC Note=kcat is 0.033 min(-1). {ECO:0000269|PubMed:20122937};
CC Temperature dependence:
CC Optimum temperature is 70 degrees Celsius.
CC {ECO:0000269|PubMed:20122937};
CC -!- SUBUNIT: Forms a ternary complex with MCM helicase and DNA. Component
CC of the archaeal exosome complex. Interacts with Csl4 but not with Rrp4.
CC {ECO:0000255|HAMAP-Rule:MF_00007, ECO:0000269|PubMed:12947419,
CC ECO:0000269|PubMed:17078816, ECO:0000269|PubMed:20122937,
CC ECO:0000269|PubMed:22503705, ECO:0000269|PubMed:23324612,
CC ECO:0000269|PubMed:23357171}.
CC -!- SIMILARITY: Belongs to the archaeal DnaG primase family.
CC {ECO:0000255|HAMAP-Rule:MF_00007}.
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DR EMBL; Y08257; CAA69522.1; -; Genomic_DNA.
DR EMBL; AE006641; AAK40440.1; -; Genomic_DNA.
DR PIR; S75408; S75408.
DR AlphaFoldDB; P95980; -.
DR SMR; P95980; -.
DR IntAct; P95980; 2.
DR MINT; P95980; -.
DR STRING; 273057.SSO0079; -.
DR PRIDE; P95980; -.
DR EnsemblBacteria; AAK40440; AAK40440; SSO0079.
DR KEGG; sso:SSO0079; -.
DR PATRIC; fig|273057.12.peg.79; -.
DR eggNOG; arCOG04281; Archaea.
DR HOGENOM; CLU_034626_0_0_2; -.
DR InParanoid; P95980; -.
DR OMA; KYMIVAQ; -.
DR PhylomeDB; P95980; -.
DR BRENDA; 2.7.7.101; 6163.
DR Proteomes; UP000001974; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR GO; GO:0000428; C:DNA-directed RNA polymerase complex; IEA:UniProtKB-KW.
DR GO; GO:0000178; C:exosome (RNase complex); IEA:UniProtKB-KW.
DR GO; GO:1990077; C:primosome complex; IEA:UniProtKB-KW.
DR GO; GO:0003896; F:DNA primase activity; IBA:GO_Central.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008143; F:poly(A) binding; IEA:InterPro.
DR GO; GO:0006269; P:DNA replication, synthesis of RNA primer; IBA:GO_Central.
DR CDD; cd01029; TOPRIM_primases; 1.
DR HAMAP; MF_00007; DNA_primase_DnaG_arc; 1.
DR InterPro; IPR020607; Primase_DnaG_arc.
DR InterPro; IPR034154; TOPRIM_DnaG/twinkle.
DR InterPro; IPR006171; TOPRIM_domain.
DR Pfam; PF13662; Toprim_4; 1.
DR SMART; SM00493; TOPRIM; 1.
DR PROSITE; PS50880; TOPRIM; 1.
PE 1: Evidence at protein level;
KW DNA replication; DNA-directed RNA polymerase; Exosome; Magnesium;
KW Metal-binding; Nucleotidyltransferase; Primosome; Reference proteome;
KW Transcription; Transferase.
FT CHAIN 1..406
FT /note="DNA primase DnaG"
FT /id="PRO_0000144132"
FT DOMAIN 169..247
FT /note="Toprim"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00007"
FT BINDING 175
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00007"
FT BINDING 220
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00007"
FT BINDING 220
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00007"
FT BINDING 222
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00007"
FT MUTAGEN 175
FT /note="E->Q: Strong decrease of primase activity."
FT /evidence="ECO:0000269|PubMed:20122937"
FT MUTAGEN 179
FT /note="D->A: Decreases binding affinity to DNA. Does not
FT affect primase activity."
FT /evidence="ECO:0000269|PubMed:23357171"
FT MUTAGEN 220
FT /note="D->A: Decreases binding affinity to DNA and primase
FT activity."
FT /evidence="ECO:0000269|PubMed:23357171"
FT MUTAGEN 222
FT /note="D->A: Decreases binding affinity to DNA and primase
FT activity."
FT /evidence="ECO:0000269|PubMed:23357171"
SQ SEQUENCE 406 AA; 45410 MW; 26E5B615560C6EF1 CRC64;
MSFQMKYDIR LRFEVEGIVE KTDVIGAIFG QTENLFGDEF DLRELQDKGR LGRIIVEVKT
KGGKSEGEII IPSNLDRIET ALIAAMVESV DKVGPYNSKF ELIEIEDIRA EKLKKIIERA
KGILSSWSKE KSLDIKEVIN EISSAVKVGE ITEYGPERLP AGPDVDKDPN LIIVEGRADV
INLLRYGYKN VIAVEGATSR IPETLINLSK MKKTVIAFLD GDHGGDLILK ELLSNNVKID
FVARAPIGRE VEELTGKEIA KALSNMMPLT QYLKKVQEAE QAIAKNVIAK EEKPIQSETT
QQVVQITLPQ NILEEIKKLP GTLEGVLYDN NWNLIEKVQV RDIIPKLEAY EDNKVAYIIF
DGVITQRLLD LASQKNIKMI IGARIGGINK RPQNVDILTF TDIISS
