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P0AE70 (MAZF_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Last modified June 11, 2014. Version 71. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (3) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
mRNA interferase MazF

EC=3.1.-.-
Alternative name(s):
Endoribonuclease MazF
Toxin MazF
Gene names
Name:mazF
Synonyms:chpA, chpAK
Ordered Locus Names:b2782, JW2753
OrganismEscherichia coli (strain K12) [Reference proteome] [HAMAP]
Taxonomic identifier83333 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia

Protein attributes

Sequence length111 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Toxic component of a toxin-antitoxin (TA) module. MazF is a sequence-specific mRNA endoribonuclease that inhibits protein synthesis and induces bacterial stasis. It is very stable, single-strand specific and cleavage is independent of the ribosome, although translation enhances cleavage for some mRNAs. Cleavage usually occurs between the first A and C of ACA sequences, yielding a 2',3'-cyclic phosphate and a free 5'-OH. The endoribonuclease activity (a toxin) is inhibited by the labile cognate antitoxin MazE. Toxicity results when the levels of MazE decrease in the cell, leading to mRNA degradation. This effect can be rescued by expression of MazE, but after 6 hours in rich medium overexpression of MazF leads to programmed cell death. MazF-mediated cell death occurs following a number of stress conditions in a relA-dependent fashion and only when cells are in log phase. Cell growth and viability are not affected when MazF and MazE are coexpressed. Programmed cell death occurs when cells are at high density and depends on the presence of MazE-MazF and a quorum sensing pentapeptide, the extracellular death factor (EDF) with sequence NNWNN, probably produced from the zwg gene product glucose-6-phosphate 1-dehydrogenase. Both MazE and MazE-MazF bind to the promoter region of the mazE-mazF operon to inhibit their transcription. Ref.5 Ref.7 Ref.9 Ref.10 Ref.11 Ref.12 Ref.18

Might also serve to protect cells against bacteriophage; in the presence of MazE-MazF fewer P1 phage are produced than in a disruption strain. For strain K38 most wild-type cells are killed but not by phage lysis; it was suggested that MazE-MazF causes P1 phage exclusion from the bacterial population. This phenomenon is strain dependent. Ref.5 Ref.7 Ref.9 Ref.10 Ref.11 Ref.12 Ref.18

Cell death governed by the MazE-MazF and DinJ-YafQ TA modules seems to play a role in biofilm formation, while MazE-MazF is also implicated in cell death in liquid media. Ref.5 Ref.7 Ref.9 Ref.10 Ref.11 Ref.12 Ref.18

Enzyme regulation

Inhibited by Mg2+. Ref.12

Subunit structure

Probably a dimer. Forms a heterohexamer composed of alternating toxin and antitoxin homodimers MazF(2)-MazE(2)-MazF2. The binding site of MazE and ssRNA or ssDNA are largely overlapping; the presence of only 1 MazE molecule inhibits mRNA endoribonuclease activity. Ref.6 Ref.20

Induction

Expressed in exponentially growing cells. Induction has been reported to occur after amino acid starvation in a ppGpp-independent fashion and to be Lon protease-dependent (Ref.8), but also to not occur after amino acid starvation and to be regulated by ppGpp (Ref.5). MazE alone and in combination with MazF, represses transcription of the mazE-mazF operon. Fis activates transcription. Part of the relA-mazE-mazF-mazG operon, there is also a second mazE-mazF specific promoter which is negatively autoregulated. Ref.5 Ref.6 Ref.8 Ref.12 Ref.14

Disruption phenotype

Decreased sensitivity to dramatic intracellular increases of ppGpp. Cells missing mazE-mazF survive high temperature, various DNA-damaging agents and H2O2 exposure better than wild-type cells. Cells missing mazE-mazF produce more P1 phage than wild-type cells, while introduction of lysogens into a growing non-lysogenic deletion culture is lethal. Cells missing mazE-mazF show reduced biofilm formation, and survive antibiotic treatment in log phase better than wild-type cells. Ref.5 Ref.8 Ref.9 Ref.11 Ref.18

Biotechnological use

Can be used to produce large quantities of a single protein if the gene coding for the protein does not contain any ACA codons. Up to 90% of bacterial cellular protein can be the target, which can be produced for up to 4 days. The system also works in eukaryotic cells. Ref.13

Sequence similarities

Belongs to the PemK/MazF family.

Caution

Strain K12 / MG1655 is deficient in both production and response to EDF, unlike strains K12 / MC4100, K12 / W3110 and K12 / K38, all of which make and respond to EDF.

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 111111mRNA interferase MazF
PRO_0000201897

Experimental info

Mutagenesis241E → A: Greatly reduces toxicity, about 10-fold less RNA cleavage activity. Ref.21

Secondary structure

........................ 111
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P0AE70 [UniParc].

Last modified December 6, 2005. Version 1.
Checksum: 1579C867DD6B96AC

FASTA11112,098
        10         20         30         40         50         60 
MVSRYVPDMG DLIWVDFDPT KGSEQAGHRP AVVLSPFMYN NKTGMCLCVP CTTQSKGYPF 

        70         80         90        100        110 
EVVLSGQERD GVALADQVKS IAWRARGATK KGTVAPEELQ LIKAKINVLI G 

« Hide

References

« Hide 'large scale' references
[1]"The nucleotide sequence and characterization of the relA gene of Escherichia coli."
Metzger S., Dror I.B., Aizenman E., Schreiber G., Toone M., Friesen J.D., Cashel M., Glaser G.
J. Biol. Chem. 263:15699-15704(1988) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], OPERON STRUCTURE.
Strain: K12.
[2]"chpA and chpB, Escherichia coli chromosomal homologs of the pem locus responsible for stable maintenance of plasmid R100."
Masuda Y., Miyakawa K., Nishimura Y., Ohtsubo E.
J. Bacteriol. 175:6850-6856(1993) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
Strain: K12 / MC1000 / ATCC 39531.
[3]"The complete genome sequence of Escherichia coli K-12."
Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y.
Science 277:1453-1462(1997) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: K12 / MG1655 / ATCC 47076.
[4]"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110."
Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.
Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[5]"An Escherichia coli chromosomal 'addiction module' regulated by guanosine 3',5'-bispyrophosphate: a model for programmed bacterial cell death."
Aizenman E., Engelberg-Kulka H., Glaser G.
Proc. Natl. Acad. Sci. U.S.A. 93:6059-6063(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION IN PROGRAMMED CELL DEATH, INTERACTION WITH MAZE, INDUCTION, DISRUPTION PHENOTYPE, OPERON STRUCTURE.
Strain: K12 / MC4100 / ATCC 35695 / DSM 6574.
[6]"The regulation of the Escherichia coli mazEF promoter involves an unusual alternating palindrome."
Marianovsky I., Aizenman E., Engelberg-Kulka H., Glaser G.
J. Biol. Chem. 276:5975-5984(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: TRANSCRIPTION REGULATION, SUBUNIT.
Strain: K12 / MC4100 / ATCC 35695 / DSM 6574.
[7]"Rapid induction and reversal of a bacteriostatic condition by controlled expression of toxins and antitoxins."
Pedersen K., Christensen S.K., Gerdes K.
Mol. Microbiol. 45:501-510(2002) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION AS A TOXIN.
Strain: K12.
[8]"Toxin-antitoxin loci as stress-response-elements: ChpAK/MazF and ChpBK cleave translated RNAs and are counteracted by tmRNA."
Christensen S.K., Pedersen K., Hansen F.G., Gerdes K.
J. Mol. Biol. 332:809-819(2003) [PubMed] [Europe PMC] [Abstract]
Cited for: RNA CLEAVAGE, INDUCTION, DISRUPTION PHENOTYPE.
Strain: K12 / MG1655 / ATCC 47076.
[9]"Escherichia coli mazEF-mediated cell death is triggered by various stressful conditions."
Hazan R., Sat B., Engelberg-Kulka H.
J. Bacteriol. 186:3663-3669(2004) [PubMed] [Europe PMC] [Abstract]
Cited for: DISRUPTION PHENOTYPE, FUNCTION IN STRESS RESPONSE, DEPENDENCE ON RELA.
Strain: K12 / K38 / S26 and K12 / MC4100 / ATCC 35695 / DSM 6574.
[10]"MazF-mediated cell death in Escherichia coli: a point of no return."
Amitai S., Yassin Y., Engelberg-Kulka H.
J. Bacteriol. 186:8295-8300(2004) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION IN PROGRAMMED CELL DEATH.
Strain: K12 / MC4100 / ATCC 35695 / DSM 6574.
[11]"Escherichia coli mazEF-mediated cell death as a defense mechanism that inhibits the spread of phage P1."
Hazan R., Engelberg-Kulka H.
Mol. Genet. Genomics 272:227-234(2004) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION IN PHAGE IMMUNITY, DISRUPTION PHENOTYPE.
Strain: K12 / JM109 / ATCC 53323, K12 / K38 / S26 and K12 / MC4100 / ATCC 35695 / DSM 6574.
[12]"Insights into the mRNA cleavage mechanism by MazF, an mRNA interferase."
Zhang Y., Zhang J., Hara H., Kato I., Inouye M.
J. Biol. Chem. 280:3143-3150(2005) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION AS AN ENDONUCLEASE, ENZYME REGULATION, SUBSTRATE SPECIFICITY, RNA-BINDING.
[13]"Single protein production in living cells facilitated by an mRNA interferase."
Suzuki M., Zhang J., Liu M., Woychik N.A., Inouye M.
Mol. Cell 18:253-261(2005) [PubMed] [Europe PMC] [Abstract]
Cited for: BIOTECHNOLOGY.
[14]"MazG -- a regulator of programmed cell death in Escherichia coli."
Gross M., Marianovsky I., Glaser G.
Mol. Microbiol. 59:590-601(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: INDUCTION, OPERON STRUCTURE.
Strain: K12 / MC4100 / ATCC 35695 / DSM 6574 and K12 / W3110 / ATCC 27325 / DSM 5911.
[15]"A linear pentapeptide is a quorum-sensing factor required for mazEF-mediated cell death in Escherichia coli."
Kolodkin-Gal I., Hazan R., Gaathon A., Carmeli S., Engelberg-Kulka H.
Science 318:652-655(2007) [PubMed] [Europe PMC] [Abstract]
Cited for: REQUIREMENT FOR EXTRACELLULAR DEATH FACTOR (EDF).
Strain: K12 / MC4100 / ATCC 35695 / DSM 6574.
[16]"The extracellular death factor: physiological and genetic factors influencing its production and response in Escherichia coli."
Kolodkin-Gal I., Engelberg-Kulka H.
J. Bacteriol. 190:3169-3175(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: EDF PRODUCTION, STRAIN DIFFERENCES IN ABILITY TO MAKE AND RESPOND TO EDF.
Strain: K12 / K38 / S26, K12 / MC4100 / ATCC 35695 / DSM 6574, K12 / MG1655 / ATCC 47076 and K12 / W3110 / ATCC 27325 / DSM 5911.
[17]"Translation affects YoeB and MazF messenger RNA interferase activities by different mechanisms."
Christensen-Dalsgaard M., Gerdes K.
Nucleic Acids Res. 36:6472-6481(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: TRANSLATION IMPROVES CLEAVAGE EFFICIENCY.
Strain: K12.
[18]"A differential effect of E. coli toxin-antitoxin systems on cell death in liquid media and biofilm formation."
Kolodkin-Gal I., Verdiger R., Shlosberg-Fedida A., Engelberg-Kulka H.
PLoS ONE 4:E6785-E6785(2009) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION IN CELL DEATH, FUNCTION IN BIOFILM FORMATION, DISRUPTION PHENOTYPE.
Strain: K12 / MC4100 / ATCC 35695 / DSM 6574.
[19]"mRNA interferases, sequence-specific endoribonucleases from the toxin-antitoxin systems."
Yamaguchi Y., Inouye M.
Prog. Mol. Biol. Transl. Sci. 85:467-500(2009) [PubMed] [Europe PMC] [Abstract]
Cited for: REVIEW.
[20]"Crystal structure of the MazE/MazF complex: molecular bases of antidote-toxin recognition."
Kamada K., Hanaoka F., Burley S.K.
Mol. Cell 11:875-884(2003) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) OF 2-111, POSSIBLE DNA-BINDING, SUBUNIT.
[21]"Characterization of dual substrate binding sites in the homodimeric structure of Escherichia coli mRNA interferase MazF."
Li G.Y., Zhang Y., Chan M.C., Mal T.K., Hoeflich K.P., Inouye M., Ikura M.
J. Mol. Biol. 357:139-150(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: STRUCTURE BY NMR, MUTAGENESIS OF GLU-24.
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
D16450 Genomic DNA. Translation: BAA03918.1.
J04039 Unassigned DNA. Translation: AAA03239.1.
U29580 Genomic DNA. Translation: AAA69292.1.
U00096 Genomic DNA. Translation: AAC75824.1.
AP009048 Genomic DNA. Translation: BAE76856.1.
PIRB49339.
RefSeqNP_417262.1. NC_000913.3.
YP_490990.1. NC_007779.1.

3D structure databases

PDBe
RCSB-PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1UB4X-ray1.70A/B2-111[»]
3NFCX-ray2.00A/B/C/D/E/F1-111[»]
DisProtDP00299.
ProteinModelPortalP0AE70.
SMRP0AE70. Positions 2-111.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

IntActP0AE70. 2 interactions.
STRING511145.b2782.

Chemistry

BindingDBP0AE70.
ChEMBLCHEMBL1795096.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaAAC75824; AAC75824; b2782.
BAE76856; BAE76856; BAE76856.
GeneID12933306.
947252.
KEGGecj:Y75_p2719.
eco:b2782.
PATRIC32120980. VBIEscCol129921_2882.

Organism-specific databases

EchoBASEEB1229.
EcoGeneEG11249. mazF.

Phylogenomic databases

eggNOGCOG2337.
HOGENOMHOG000290185.
KOK07171.
OMAQAGREQS.
OrthoDBEOG67X204.
PhylomeDBP0AE70.

Enzyme and pathway databases

BioCycEcoCyc:EG11249-MONOMER.
ECOL316407:JW2753-MONOMER.

Gene expression databases

GenevestigatorP0AE70.

Family and domain databases

Gene3D2.30.30.110. 1 hit.
InterProIPR003477. PemK.
IPR011067. Plasmid_toxin/cell-grow_inhib.
[Graphical view]
PfamPF02452. PemK. 1 hit.
[Graphical view]
SUPFAMSSF50118. SSF50118. 1 hit.
ProtoNetSearch...

Other

EvolutionaryTraceP0AE70.
PROP0AE70.

Entry information

Entry nameMAZF_ECOLI
AccessionPrimary (citable) accession number: P0AE70
Secondary accession number(s): P33645, Q2MA50
Entry history
Integrated into UniProtKB/Swiss-Prot: December 6, 2005
Last sequence update: December 6, 2005
Last modified: June 11, 2014
This is version 71 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

Escherichia coli

Escherichia coli (strain K12): entries and cross-references to EcoGene