TRAM1_ECOLI
ID TRAM1_ECOLI Reviewed; 127 AA.
AC P10026;
DT 01-JUL-1989, integrated into UniProtKB/Swiss-Prot.
DT 01-JUL-1989, sequence version 1.
DT 03-AUG-2022, entry version 128.
DE RecName: Full=Relaxosome protein TraM;
GN Name=traM; OrderedLocusNames=ECOK12F071;
OS Escherichia coli (strain K12).
OG Plasmid F.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC PLASMID=F;
RX PubMed=6298579; DOI=10.1007/bf00330058;
RA Thompson R., Taylor L.;
RT "Promoter mapping and DNA sequencing of the F plasmid transfer genes traM
RT and traJ.";
RL Mol. Gen. Genet. 188:513-518(1982).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC PLASMID=F;
RX PubMed=7915817; DOI=10.1128/mr.58.2.162-210.1994;
RA Frost L.S., Ippen-Ihler K., Skurray R.A.;
RT "Analysis of the sequence and gene products of the transfer region of the F
RT sex factor.";
RL Microbiol. Rev. 58:162-210(1994).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / CR63; PLASMID=F;
RA Shimizu H., Saitoh Y., Suda Y., Uehara K., Sampei G., Mizobuchi K.;
RT "Complete nucleotide sequence of the F plasmid: its implications for
RT organization and diversification of plasmid genomes.";
RL Submitted (APR-2000) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP FUNCTION IN DNA-BINDING, AND SUBCELLULAR LOCATION.
RC PLASMID=F;
RX PubMed=1479887; DOI=10.1111/j.1365-2958.1992.tb01754.x;
RA Di Laurenzio L., Frost L.S., Paranchych W.;
RT "The TraM protein of the conjugative plasmid F binds to the origin of
RT transfer of the F and ColE1 plasmids.";
RL Mol. Microbiol. 6:2951-2959(1992).
RN [5]
RP INDUCTION, FUNCTION IN AUTOREGULATION, AND DISRUPTION PHENOTYPE.
RC PLASMID=F;
RX PubMed=8736534; DOI=10.1046/j.1365-2958.1996.5361059.x;
RA Penfold S.S., Simon J., Frost L.S.;
RT "Regulation of the expression of the traM gene of the F sex factor of
RT Escherichia coli.";
RL Mol. Microbiol. 20:549-558(1996).
RN [6]
RP INTERACTION WITH TRAD.
RC PLASMID=F;
RX PubMed=9324263; DOI=10.1128/jb.179.19.6133-6137.1997;
RA Disque-Kochem C., Dreiseikelmann B.;
RT "The cytoplasmic DNA-binding protein TraM binds to the inner membrane
RT protein TraD in vitro.";
RL J. Bacteriol. 179:6133-6137(1997).
RN [7]
RP FUNCTION IN COOPERATIVITY OF DNA-BINDING, AND FUNCTION IN DNA-BENDING.
RC PLASMID=F;
RX PubMed=11875064; DOI=10.1074/jbc.m111682200;
RA Fekete R.A., Frost L.S.;
RT "Characterizing the DNA contacts and cooperative binding of F plasmid TraM
RT to its cognate sites at oriT.";
RL J. Biol. Chem. 277:16705-16711(2002).
RN [8]
RP INTERACTION WITH TRAD, AND MUTAGENESIS OF LYS-99 AND PHE-121.
RC PLASMID=F;
RX PubMed=15995191; DOI=10.1128/jb.187.14.4767-4773.2005;
RA Lu J., Frost L.S.;
RT "Mutations in the C-terminal region of TraM provide evidence for in vivo
RT TraM-TraD interactions during F-plasmid conjugation.";
RL J. Bacteriol. 187:4767-4773(2005).
RN [9]
RP FUNCTION IN STIMULATING TRAI NICKING, DNA-BINDING, INTERACTION WITH TRAY,
RP AND SUBUNIT.
RC PLASMID=F;
RX PubMed=17238924; DOI=10.1111/j.1365-2958.2006.05576.x;
RA Ragonese H., Haisch D., Villareal E., Choi J.H., Matson S.W.;
RT "The F plasmid-encoded TraM protein stimulates relaxosome-mediated cleavage
RT at oriT through an interaction with TraI.";
RL Mol. Microbiol. 63:1173-1184(2007).
RN [10]
RP SUBUNIT.
RC PLASMID=F;
RX PubMed=21565799; DOI=10.1093/nar/gkr296;
RA Wong J.J., Lu J., Edwards R.A., Frost L.S., Glover J.N.;
RT "Structural basis of cooperative DNA recognition by the plasmid conjugation
RT factor, TraM.";
RL Nucleic Acids Res. 39:6775-6788(2011).
RN [11]
RP X-RAY CRYSTALLOGRAPHY (1.4 ANGSTROMS) OF 58-127, COILED-COIL DOMAIN,
RP SUBUNIT, DNA-BINDING, AND MUTAGENESIS OF GLU-88.
RC PLASMID=F;
RX PubMed=16710295; DOI=10.1038/sj.emboj.7601151;
RA Lu J., Edwards R.A., Wong J.J., Manchak J., Scott P.G., Frost L.S.,
RA Glover J.N.;
RT "Protonation-mediated structural flexibility in the F conjugation
RT regulatory protein, TraM.";
RL EMBO J. 25:2930-2939(2006).
RN [12]
RP X-RAY CRYSTALLOGRAPHY (2.55 ANGSTROMS) OF 58-127 IN COMPLEX WITH C-TERMINUS
RP OF TRAD, AND MUTAGENESIS OF ASN-5; LYS-76; LYS-99; VAL-106 AND ARG-110.
RC PLASMID=F;
RX PubMed=18717787; DOI=10.1111/j.1365-2958.2008.06391.x;
RA Lu J., Wong J.J., Edwards R.A., Manchak J., Frost L.S., Glover J.N.;
RT "Structural basis of specific TraD-TraM recognition during F plasmid-
RT mediated bacterial conjugation.";
RL Mol. Microbiol. 70:89-99(2008).
CC -!- FUNCTION: Conjugative DNA transfer (CDT) is the unidirectional transfer
CC of ssDNA plasmid from a donor to a recipient cell. It is the central
CC mechanism by which antibiotic resistance and virulence factors are
CC propagated in bacterial populations. Part of the relaxosome, which
CC facilitates a site- and strand-specific cut in the origin of transfer
CC by TraI, at the nic site. Cooperatively binds 3 regions in the F
CC plasmid oriT locus; 2 are required for autoregulation while the other
CC is required for plasmid transfer. Bends oriT DNA less than 50 degrees.
CC Plasmid specificity is conferred by the TraD-TraM pair.
CC {ECO:0000269|PubMed:11875064, ECO:0000269|PubMed:1479887,
CC ECO:0000269|PubMed:17238924, ECO:0000269|PubMed:8736534}.
CC -!- SUBUNIT: Homotetramer. 2 homotetramers cooperatively bind to DNA
CC although they do not contact each other; cooperativity is achieved by
CC DNA kinking and unwinding (Probable). Part of the relaxosome, a complex
CC composed of plasmid encoded TraI, TraM, TraY and host-encoded IHF which
CC binds to the F plasmid origin of transfer (oriT) in a site- and
CC sequence-specific manner. Interacts with TraD. Also interacts with
CC TraY. {ECO:0000269|PubMed:15995191, ECO:0000269|PubMed:16710295,
CC ECO:0000269|PubMed:17238924, ECO:0000269|PubMed:18717787,
CC ECO:0000269|PubMed:21565799, ECO:0000269|PubMed:9324263, ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:1479887}.
CC -!- INDUCTION: Regulates its own expression from two promoters, requires
CC TraY as well as expression of the tra operon for maximal transcription.
CC TraM is in a monocistonic operon. {ECO:0000269|PubMed:8736534}.
CC -!- DISRUPTION PHENOTYPE: Loss of conjugative DNA transfer.
CC {ECO:0000269|PubMed:8736534}.
CC -!- SIMILARITY: Belongs to the relaxosome TraM family. {ECO:0000305}.
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DR EMBL; K01147; AAA24907.1; -; Genomic_DNA.
DR EMBL; X00545; CAA25216.1; -; Genomic_DNA.
DR EMBL; U01159; AAC44191.1; -; Genomic_DNA.
DR EMBL; AP001918; BAA97941.1; -; Genomic_DNA.
DR PIR; A29331; JCECMF.
DR RefSeq; NP_061450.1; NC_002483.1.
DR RefSeq; WP_001151527.1; NZ_CP014273.1.
DR PDB; 2G7O; X-ray; 1.40 A; A=58-127.
DR PDB; 2G9E; X-ray; 1.80 A; A=58-127.
DR PDB; 3D8A; X-ray; 2.55 A; A/B/C/D/E/F/G/H=58-127.
DR PDB; 4QPO; X-ray; 2.00 A; A/B/C/D=2-54.
DR PDB; 4QPQ; X-ray; 3.11 A; A/B/C/D/E/F/G/H=2-54.
DR PDBsum; 2G7O; -.
DR PDBsum; 2G9E; -.
DR PDBsum; 3D8A; -.
DR PDBsum; 4QPO; -.
DR PDBsum; 4QPQ; -.
DR AlphaFoldDB; P10026; -.
DR SMR; P10026; -.
DR DIP; DIP-27653N; -.
DR PRIDE; P10026; -.
DR PATRIC; fig|83333.107.peg.642; -.
DR EvolutionaryTrace; P10026; -.
DR PRO; PR:P10026; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR CDD; cd14804; Tra_M; 1.
DR Gene3D; 1.10.10.450; -; 1.
DR InterPro; IPR010992; IHF-like_DNA-bd_dom_sf.
DR InterPro; IPR042073; TraM_DNA-bd.
DR InterPro; IPR007925; TRelaxosome_TraM.
DR Pfam; PF05261; Tra_M; 1.
DR SUPFAM; SSF47729; SSF47729; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Coiled coil; Conjugation; Cytoplasm; DNA-binding; Plasmid;
KW Repressor; Transcription; Transcription regulation.
FT CHAIN 1..127
FT /note="Relaxosome protein TraM"
FT /id="PRO_0000068467"
FT COILED 63..89
FT /evidence="ECO:0000269|PubMed:16710295"
FT MUTAGEN 5
FT /note="N->D: Loss of all functions."
FT /evidence="ECO:0000269|PubMed:18717787"
FT MUTAGEN 76
FT /note="K->E: 100,000-fold decrease in conjugation
FT efficiency, but tetramerizes and binds DNA normally."
FT /evidence="ECO:0000269|PubMed:18717787"
FT MUTAGEN 88
FT /note="E->L,Q: Increased homotetramer stability."
FT /evidence="ECO:0000269|PubMed:16710295"
FT MUTAGEN 99
FT /note="K->E: 100,000-fold decrease in conjugation
FT efficiency, but tetramerizes and binds DNA normally. Binds
FT less well to TraD. May be dominant over wild-type.
FT Partially rescued by a TraD E-712 mutation."
FT /evidence="ECO:0000269|PubMed:15995191,
FT ECO:0000269|PubMed:18717787"
FT MUTAGEN 106
FT /note="V->A: 2000-fold decrease in conjugation efficiency,
FT but tetramerizes and binds DNA normally."
FT /evidence="ECO:0000269|PubMed:18717787"
FT MUTAGEN 110
FT /note="R->E: 33,000-fold decrease in conjugation
FT efficiency, but tetramerize and bind DNA normally."
FT /evidence="ECO:0000269|PubMed:18717787"
FT MUTAGEN 121
FT /note="F->S: Alters oligomerization, probably more dimers
FT than tetramers."
FT /evidence="ECO:0000269|PubMed:15995191"
FT STRAND 3..7
FT /evidence="ECO:0007829|PDB:4QPO"
FT HELIX 10..24
FT /evidence="ECO:0007829|PDB:4QPO"
FT STRAND 30..33
FT /evidence="ECO:0007829|PDB:4QPO"
FT HELIX 35..53
FT /evidence="ECO:0007829|PDB:4QPO"
FT HELIX 63..89
FT /evidence="ECO:0007829|PDB:2G7O"
FT HELIX 92..94
FT /evidence="ECO:0007829|PDB:2G7O"
FT HELIX 98..100
FT /evidence="ECO:0007829|PDB:2G7O"
FT HELIX 102..120
FT /evidence="ECO:0007829|PDB:2G7O"
FT STRAND 123..125
FT /evidence="ECO:0007829|PDB:2G9E"
SQ SEQUENCE 127 AA; 14507 MW; FC4642125B7E7CAC CRC64;
MAKVNLYISN DAYEKINAII EKRRQEGARE KDVSFSATAS MLLELGLRVH EAQMERKESA
FNQTEFNKLL LECVVKTQSS VAKILGIESL SPHVSGNSKF EYANMVEDIR EKVSSEMERF
FPKNDDE
