MOBA2_ECOLX
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 03-AUG-2022, entry version 114.
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; Proteobacteria; 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 RefSeq; WP_001395566.1; NZ_VTMX01000009.1.
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 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
