ID   MOBA2_ECOLX             Reviewed;         709 AA.
AC   P07112; Q60198;
DT   01-APR-1988, integrated into UniProtKB/Swiss-Prot.
DT   23-JAN-2007, sequence version 4.
DT   28-JUN-2023, entry version 118.
DE   RecName: Full=Mobilization protein A;
DE   AltName: Full=DNA strand transferase;
DE   Includes:
DE     RecName: Full=DNA relaxase;
DE              EC=5.6.2.1;
DE     AltName: Full=DNA nickase;
DE   Includes:
DE     RecName: Full=DNA primase;
DE              EC=2.7.7.101 {ECO:0000269|PubMed:10217797};
GN   Name=mobA; Synonyms=repB;
OS   Escherichia coli.
OG   Plasmid IncQ RSF1010, and Plasmid R1162.
OC   Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales;
OC   Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=562;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   PLASMID=IncQ RSF1010;
RX   PubMed=2653965; DOI=10.1016/0378-1119(89)90273-4;
RA   Scholz P., Haring V., Wittmann-Liebold B., Ashman K., Bagdasarian M.,
RA   Scherzinger E.;
RT   "Complete nucleotide sequence and gene organization of the broad-host-range
RT   plasmid RSF1010.";
RL   Gene 75:271-288(1989).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-407.
RC   PLASMID=IncQ RSF1010;
RX   PubMed=3033438; DOI=10.1007/bf00326552;
RA   Derbyshire K.M., Hatfull G., Willetts N.;
RT   "Mobilization of the non-conjugative plasmid RSF1010: a genetic and DNA
RT   sequence analysis of the mobilization region.";
RL   Mol. Gen. Genet. 206:161-168(1987).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-400.
RC   PLASMID=IncQ RSF1010;
RX   PubMed=1563624; DOI=10.1016/0378-1119(92)90675-f;
RA   Frey J., Bagdasarian M.M., Bagdasarian M.;
RT   "Replication and copy number control of the broad-host-range plasmid
RT   RSF1010.";
RL   Gene 113:101-106(1992).
RN   [4]
RP   PROTEIN SEQUENCE OF 2-15 AND 23-31, AND COFACTOR.
RC   PLASMID=IncQ RSF1010;
RX   PubMed=8223650; DOI=10.1111/j.1432-1033.1993.tb18323.x;
RA   Scherzinger E., Kruft V., Otto S.;
RT   "Purification of the large mobilization protein of plasmid RSF1010 and
RT   characterization of its site-specific DNA-cleaving/DNA-joining activity.";
RL   Eur. J. Biochem. 217:929-938(1993).
RN   [5]
RP   FUNCTION IN DNA CLEAVAGE, AND COFACTOR.
RC   PLASMID=IncQ RSF1010;
RX   PubMed=1738602; DOI=10.1093/nar/20.1.41;
RA   Scherzinger E., Lurz R., Otto S., Dobrinski B.;
RT   "In vitro cleavage of double- and single-stranded DNA by plasmid RSF1010-
RT   encoded mobilization proteins.";
RL   Nucleic Acids Res. 20:41-48(1992).
RN   [6]
RP   BINDING TO SINGLE-STRANDED ORIT.
RC   PLASMID=R1162;
RX   PubMed=8233790; DOI=10.1093/nar/21.19.4563;
RA   Bhattacharjee M.K., Meyer R.J.;
RT   "Specific binding of MobA, a plasmid-encoded protein involved in the
RT   initiation and termination of conjugal DNA transfer, to single-stranded
RT   oriT DNA.";
RL   Nucleic Acids Res. 21:4563-4568(1993).
RN   [7]
RP   FUNCTION OF PRIMASE DOMAIN IN CONJUGAL TRANSFER.
RC   PLASMID=R1162;
RX   PubMed=8955311; DOI=10.1128/jb.178.23.6888-6894.1996;
RA   Henderson D., Meyer R.J.;
RT   "The primase of broad-host-range plasmid R1162 is active in conjugal
RT   transfer.";
RL   J. Bacteriol. 178:6888-6894(1996).
RN   [8]
RP   PRIMASE ACTIVITY, AND CATALYTIC ACTIVITY.
RC   PLASMID=R1162;
RX   PubMed=10217797; DOI=10.1128/jb.181.9.2973-2978.1999;
RA   Henderson D., Meyer R.;
RT   "The MobA-linked primase is the only replication protein of R1162 required
RT   for conjugal mobilization.";
RL   J. Bacteriol. 181:2973-2978(1999).
RN   [9]
RP   TRANSFERASE ACTIVITY.
RC   PLASMID=R1162;
RX   PubMed=11839744; DOI=10.1074/jbc.m110759200;
RA   Becker E.C., Meyer R.J.;
RT   "MobA, the DNA strand transferase of plasmid R1162: the minimal domain
RT   required for DNA processing at the origin of transfer.";
RL   J. Biol. Chem. 277:14575-14580(2002).
RN   [10]
RP   SPECIFICITY OF DNA SEQUENCE FOR CLEAVAGE.
RC   PLASMID=R1162;
RX   PubMed=12775691; DOI=10.1128/jb.185.12.3538-3546.2003;
RA   Becker E.C., Meyer R.J.;
RT   "Relaxed specificity of the R1162 nickase: a model for evolution of a
RT   system for conjugative mobilization of plasmids.";
RL   J. Bacteriol. 185:3538-3546(2003).
RN   [11]
RP   X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF 2-186 IN COMPLEX WITH MANGANESE,
RP   REACTION MECHANISM, AND MUTAGENESIS OF TYR-25; TYR-32; GLU-38; GLU-74 AND
RP   GLU-76.
RC   PLASMID=R1162;
RX   PubMed=17157875; DOI=10.1016/j.jmb.2006.11.031;
RA   Monzingo A.F., Ozburn A., Xia S., Meyer R.J., Robertus J.D.;
RT   "The structure of the minimal relaxase domain of MobA at 2.1 A
RT   resolution.";
RL   J. Mol. Biol. 366:165-178(2007).
CC   -!- FUNCTION: Part of the relaxosome complex that is responsible for
CC       plasmid transfer during conjugation. Locally unwinds DNA and catalyzes
CC       the cleavage of one of the DNA strands at oriT. The cleaved strand is
CC       then transferred through the dedicated type IV secretion apparatus.
CC       MobA remains covalently linked at the 5' end of the strand, and once in
CC       the recipient cell, it probably catalyzes the rejoining of the two ends
CC       of the strand, re-forming the circular plasmid DNA. The primase
CC       activity of MobA is essential for the synthesis of primers that will
CC       initiate the DNA replication events necessary to form the double-
CC       stranded plasmid in the recipient cell. {ECO:0000269|PubMed:1738602,
CC       ECO:0000269|PubMed:8955311}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=ATP-independent breakage of single-stranded DNA, followed by
CC         passage and rejoining.; EC=5.6.2.1;
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:0000269|PubMed:10217797};
CC   -!- COFACTOR:
CC       Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC         Evidence={ECO:0000269|PubMed:1738602, ECO:0000269|PubMed:8223650};
CC       Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC         Evidence={ECO:0000269|PubMed:1738602, ECO:0000269|PubMed:8223650};
CC       Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC         Evidence={ECO:0000269|PubMed:1738602, ECO:0000269|PubMed:8223650};
CC       Name=Ba(2+); Xref=ChEBI:CHEBI:37136;
CC         Evidence={ECO:0000269|PubMed:1738602, ECO:0000269|PubMed:8223650};
CC       Note=Divalent metal cation. Can use Mg(2+), or to a lesser extent,
CC       Mn(2+), Ca(2+) or Ba(2+). {ECO:0000269|PubMed:1738602,
CC       ECO:0000269|PubMed:8223650};
CC   -!- SUBUNIT: Interacts with MobB and MobC to form the relaxosome.
CC       {ECO:0000269|PubMed:17157875}.
CC   -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC   -!- SIMILARITY: Belongs to the MobA/MobL family. {ECO:0000305}.
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DR   EMBL; M28829; AAA26445.1; -; Genomic_DNA.
DR   EMBL; X04830; CAA28520.1; -; Genomic_DNA.
DR   EMBL; S96966; AAB22064.1; -; Genomic_DNA.
DR   PIR; JH0126; JH0126.
DR   RefSeq; NP_044304.1; NC_001740.1.
DR   PDB; 2NS6; X-ray; 2.10 A; A=2-186.
DR   PDBsum; 2NS6; -.
DR   AlphaFoldDB; P07112; -.
DR   SMR; P07112; -.
DR   IntAct; P07112; 1.
DR   BindingDB; P07112; -.
DR   ChEMBL; CHEMBL5420; -.
DR   EvolutionaryTrace; P07112; -.
DR   PRO; PR:P07112; -.
DR   GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR   GO; GO:0000428; C:DNA-directed RNA polymerase complex; IEA:UniProtKB-KW.
DR   GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR   GO; GO:0003896; F:DNA primase activity; IEA:UniProtKB-EC.
DR   GO; GO:0003917; F:DNA topoisomerase type I (single strand cut, ATP-independent) activity; IEA:UniProtKB-EC.
DR   GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR   Gene3D; 1.10.1240.50; -; 1.
DR   Gene3D; 3.30.930.30; -; 1.
DR   Gene3D; 3.30.1490.240; RepB DNA-primase, N-terminal domain; 1.
DR   Gene3D; 3.30.70.1790; RepB DNA-primase, N-terminal domain; 1.
DR   Gene3D; 1.20.5.460; Single helix bin; 1.
DR   InterPro; IPR005053; MobA_MobL.
DR   InterPro; IPR039459; RepB-like_DNA_primase_dom.
DR   Pfam; PF03389; MobA_MobL; 1.
DR   Pfam; PF16793; RepB_primase; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Calcium; Coiled coil; Conjugation; Cytoplasm;
KW   Direct protein sequencing; DNA-binding; DNA-directed RNA polymerase;
KW   Isomerase; Magnesium; Manganese; Metal-binding; Mobility protein;
KW   Multifunctional enzyme; Plasmid; Topoisomerase; Transcription; Transferase.
FT   INIT_MET        1
FT                   /note="Removed"
FT                   /evidence="ECO:0000269|PubMed:8223650"
FT   CHAIN           2..709
FT                   /note="Mobilization protein A"
FT                   /id="PRO_0000210849"
FT   REGION          1..21
FT                   /note="Disordered"
FT                   /evidence="ECO:0000256|SAM:MobiDB-lite"
FT   REGION          2..186
FT                   /note="DNA relaxase"
FT   REGION          251..337
FT                   /note="Disordered"
FT                   /evidence="ECO:0000256|SAM:MobiDB-lite"
FT   REGION          315..709
FT                   /note="DNA primase"
FT   REGION          689..709
FT                   /note="Disordered"
FT                   /evidence="ECO:0000256|SAM:MobiDB-lite"
FT   COILED          567..594
FT                   /evidence="ECO:0000255"
FT   COMPBIAS        251..285
FT                   /note="Basic and acidic residues"
FT                   /evidence="ECO:0000256|SAM:MobiDB-lite"
FT   COMPBIAS        323..337
FT                   /note="Basic and acidic residues"
FT                   /evidence="ECO:0000256|SAM:MobiDB-lite"
FT   ACT_SITE        25
FT                   /note="O-(5'-phospho-DNA)-tyrosine intermediate; for
FT                   relaxase activity"
FT   BINDING         112
FT                   /ligand="a divalent metal cation"
FT                   /ligand_id="ChEBI:CHEBI:60240"
FT   BINDING         120
FT                   /ligand="a divalent metal cation"
FT                   /ligand_id="ChEBI:CHEBI:60240"
FT   BINDING         122
FT                   /ligand="a divalent metal cation"
FT                   /ligand_id="ChEBI:CHEBI:60240"
FT   SITE            70
FT                   /note="Involved in DNA binding"
FT                   /evidence="ECO:0000305"
FT   MUTAGEN         25
FT                   /note="Y->F: 99.5% decrease in conjugal transfer
FT                   frequency."
FT                   /evidence="ECO:0000269|PubMed:17157875"
FT   MUTAGEN         32
FT                   /note="Y->F: No decrease in conjugal transfer frequency."
FT                   /evidence="ECO:0000269|PubMed:17157875"
FT   MUTAGEN         38
FT                   /note="E->A: No decrease in conjugal transfer frequency."
FT                   /evidence="ECO:0000269|PubMed:17157875"
FT   MUTAGEN         74
FT                   /note="E->A: 50% decrease in conjugal transfer frequency.
FT                   10-fold decrease in conjugal transfer frequency and great
FT                   reduction in the rate of DNA cleavage; when associated with
FT                   A-76."
FT                   /evidence="ECO:0000269|PubMed:17157875"
FT   MUTAGEN         74
FT                   /note="E->Q: 70% decrease in conjugal transfer frequency."
FT                   /evidence="ECO:0000269|PubMed:17157875"
FT   MUTAGEN         76
FT                   /note="E->A: No decrease in conjugal transfer frequency.
FT                   10-fold decrease in conjugal transfer frequency and great
FT                   reduction in the rate of DNA cleavage; when associated with
FT                   A-74."
FT                   /evidence="ECO:0000269|PubMed:17157875"
FT   CONFLICT        291
FT                   /note="P -> Q (in Ref. 3; AAB22064)"
FT                   /evidence="ECO:0000305"
SQ   SEQUENCE   709 AA;  77952 MW;  DDEACDC3CA53D3E1 CRC64;
     MAIYHLTAKT GSRSGGQSAR AKADYIQREG KYARDMDEVL HAESGHMPEF VERPADYWDA
     ADLYERANGR LFKEVEFALP VELTLDQQKA LASEFAQHLT GAERLPYTLA IHAGGGENPH
     CHLMISERIN DGIERPAAQW FKRYNGKTPE KGGAQKTEAL KPKAWLEQTR EAWADHANRA
     LERAGHDARI DHRTLEAQGI ERLPGVHLGP NVVEMEGRGI RTDRADVALN IDTANAQIID
     LQEYREAIDH ERNRQSEEIQ RHQRVSGADR TAGPEHGDTG RRSPAGHEPD PAGQRGAGGG
     VAESPAPDRG GMGGAGQRVA GGSRRGEQRR AERPERVAGV ALEAMANRDA GFHDAYGGAA
     DRIVALARPD ATDNRGRLDL AALGGPMKND RTLQAIGRQL KAMGCERFDI GVRDATTGQM
     MNREWSAAEV LQNTPWLKRM NAQGNDVYIR PAEQERHGLV LVDDLSEFDL DDMKAEGREP
     ALVVETSPKN YQAWVKVADA AGGELRGQIA RTLASEYDAD PASADSRHYG RLAGFTNRKD
     KHTTRAGYQP WVLLRESKGK TATAGPALVQ QAGQQIEQAQ RQQEKARRLA SLELPERQLS
     RHRRTALDEY RSEMAGLVKR FGDDLSKCDF IAAQKLASRG RSAEEIGKAM AEASPALAER
     KPGHEADYIE RTVSKVMGLP SVQLARAELA RAPAPRQRGM DRGGPDFSM
//