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P13658 (MBEA_ECOLX) Reviewed, UniProtKB/Swiss-Prot

Last modified June 11, 2014. Version 48. Feed History...

Clusters with 100%, 90%, 50% identity | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Interactions·Sequence annotation·Sequences·References·Cross-refs·Entry infoCustomize order

Names and origin

Protein namesRecommended name:
DNA relaxase MbeA

EC=5.99.1.2
Alternative name(s):
DNA nickase
Mobilization protein MbeA
Gene names
Name:mbeA
Encoded onPlasmid ColE1
OrganismEscherichia coli
Taxonomic identifier562 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia

Protein attributes

Sequence length517 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Relaxase involved in plasmid ColE1 conjugative mobilization and is thus essential to promote the specific transfer of the plasmid during conjugation. First catalyzes the specific cleavage of one of the DNA strands at oriT, forming a covalent 5'-phosphotyrosine intermediate. The nic site corresponds to 5'-(1469)CTGG/CTTA(1462)-3' in the cleaved strand. The cleaved strand is then transferred through the dedicated type IV secretion apparatus. MbeA remains covalently linked at the 5' end of the strand, and once in the recipient cell, it probably catalyzes the rejoining of the two ends of the strand, re-forming the circular plasmid DNA. Is functional in vitro without a requirement for the conjugative accessory proteins. Ref.2

Catalytic activity

ATP-independent breakage of single-stranded DNA, followed by passage and rejoining.

Cofactor

Divalent metal cation. Can use Mg2+, Co2+ and Ni2+ with similar efficiencies, but is not active in the presence of Mn2+. Ref.2

Subunit structure

Interacts with MbeB and MbeC to form the relaxosome.

Miscellaneous

The 100 C-terminal amino acids are dispensable for activity.

Sequence similarities

To E.coli MbaA and MbkA.

Ontologies

Binary interactions

With

Entry

#Exp.

IntAct

Notes

mbeCP136572EBI-7798346,EBI-7798318

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 517517DNA relaxase MbeA
PRO_0000068400

Sites

Active site191O-(5'-phospho-DNA)-tyrosine intermediate
Metal binding971Divalent metal cation Probable
Metal binding1041Divalent metal cation Probable
Metal binding1061Divalent metal cation Probable

Experimental info

Mutagenesis191Y → A: Loss of in vitro DNA-cleavage activity and in vivo functionality. Ref.2
Mutagenesis971H → A: Loss of in vitro DNA-cleavage activity and in vivo functionality. No effect on DNA binding around the nic site. Ref.2
Mutagenesis1041E → A: Loss of in vitro DNA-cleavage activity and in vivo functionality. No effect on DNA binding around the nic site. Ref.2
Mutagenesis1041E → H: Loss of the in vitro DNA-cleavage and strand-transfer activities but displays 1% of the in vivo wild-type activity; when associated with H-106. Ref.2
Mutagenesis1061N → A: 4-fold and 3-fold reduction in the in vitro DNA-cleavage and strand-transfer activities, respectively, but no reduction in the in vivo functionality. Ref.2
Mutagenesis1061N → H: Loss of the in vitro DNA-cleavage and strand-transfer activities but displays 1% of the in vivo wild-type activity; when associated with H-104. Ref.2

Sequences

Sequence LengthMass (Da)Tools
P13658 [UniParc].

Last modified January 1, 1990. Version 1.
Checksum: 278E41B2D3B2A08A

FASTA51757,808
        10         20         30         40         50         60 
MIVKFHARGK GGGSGPVDYL LGRERNREGA TVLQGNPEEV RELIDATPFA KKYTSGVLSF 

        70         80         90        100        110        120 
AEKELPPGGR EKVMASFERV LMPGLEKNQY SILWVEHQDK GRLELNFVIP NMELQTGKRL 

       130        140        150        160        170        180 
QPYYDRADRP RIDAWQTLVN HHYGLHDPNA PENRRTLTLP DNLPETKQAL AEGVTRGIDA 

       190        200        210        220        230        240 
LYHAGEIKGR QDVIQALTEA GLEVVRVTRT SISIADPNGG KNIRLKGAFY EQSFADGRGV 

       250        260        270        280        290        300 
REKAERESRI YRENAEQRVQ EARRICKRGC DIKRDENQRR YSPVHSLDRG IAGKTPGRGE 

       310        320        330        340        350        360 
RGDDAAQEGR VKAGREYGHD VTGDSLSPVY REWRDALVSW REDTGEPGRN QEAGRDIAET 

       370        380        390        400        410        420 
EREDMGRGVC AGREQEIPCP SVREISGGDS LSGERVGTSE GVTQSDRAGN TFAERLRAAA 

       430        440        450        460        470        480 
TGLYAAAERM GERLRGIAED VFAYATGQRD AERAGHAVES AGAALERADR TLEPVIQREL 

       490        500        510 
EIREERLIQE REHVLSLERE RQPEIQERTL DGPSLGW 

« Hide

References

[1]"Characterization of the ColE1 mobilization region and its protein products."
Boyd A.C., Archer J.A.K., Sherratt D.J.
Mol. Gen. Genet. 217:488-498(1989) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]"Genetic and biochemical characterization of MbeA, the relaxase involved in plasmid ColE1 conjugative mobilization."
Varsaki A., Lucas M., Afendra A.S., Drainas C., de la Cruz F.
Mol. Microbiol. 48:481-493(2003) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION AS A RELAXASE, COFACTOR, DETERMINATION OF THE NIC SITE, MUTAGENESIS OF TYR-19; HIS-97; GLU-104 AND ASN-106.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
X15873 Genomic DNA. Translation: CAA33883.1.
PIRJQ0390.

3D structure databases

ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

IntActP13658. 1 interaction.
MINTMINT-8306165.

Proteomic databases

PRIDEP13658.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Family and domain databases

InterProIPR005094. Endonuclease_MobA/VirD2.
[Graphical view]
PfamPF03432. Relaxase. 1 hit.
[Graphical view]
ProtoNetSearch...

Entry information

Entry nameMBEA_ECOLX
AccessionPrimary (citable) accession number: P13658
Entry history
Integrated into UniProtKB/Swiss-Prot: January 1, 1990
Last sequence update: January 1, 1990
Last modified: June 11, 2014
This is version 48 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program