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Protein

Endoribonuclease MazF

Gene

mazF

Organism
Escherichia coli (strain K12)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Toxic component of a type II toxin-antitoxin (TA) module. A sequence-specific endoribonuclease it inhibits protein synthesis by cleaving mRNA and inducing bacterial stasis. It is stable, single-strand specific with mRNA cleavage independent of the ribosome, although translation enhances cleavage for some mRNAs (PubMed:18854355). Cleavage occurs at the 5'-end of ACA sequences, yielding a 2',3'-cyclic phosphate and a free 5'-OH, although cleavage can also occur on the 3'-end of the first A (PubMed:15537630, PubMed:23280569). Digests 16S rRNA in vivo 43 nts upstream of the C-terminus; this removes the anti-Shine-Dalgarno sequence forming a mixed population of wild-type and "stress ribosomes". Stress ribosomes do not translate leader-containing mRNA but are proficient in translation of leaderless mRNA, which alters the protein expression profile of the cell; MazF produces some leaderless mRNA (PubMed:21944167). The toxic endoribonuclease activity is inhibited by its 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 (PubMed:8650219, PubMed:11222603). MazF-mediated cell death occurs following a number of stress conditions in a relA-dependent fashion and only when cells are in log phase; sigma factor S (rpoS) protects stationary phase cells from MazF-killing (PubMed:15150257, PubMed:19251848). Cell growth and viability are not affected when MazF and MazE are coexpressed. Both MazE and MazE-MazF bind to the promoter region of the mazE-mazF operon to inhibit their own transcription. MazE has higher affinity for promoter DNA in the presence of MazF (PubMed:25564525). Cross-talk can occur between different TA modules. Ectopic expression of this toxin induces transcription of the relBEF TA module operon with specific cleavage of the mRNA produced (PubMed:23432955).11 Publications
Might also serve to protect cells against bacteriophage; in the presence of MazE-MazF fewer P1 phage are produced than in a disrupted 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.1 Publication
The physiological role of this TA module is debated. Programmed cell death (PCD) 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 Asn-Asn-Trp-Asn-Asn (NNWNN), probably produced from the zwf gene product glucose-6-phosphate 1-dehydrogenase (PubMed:17962566, PubMed:18310334). 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 (PubMed:19707553). Implicated in hydroxy radical-mediated cell death induced by hydroxyurea treatment (PubMed:20005847, PubMed:23416055). In conjunction with EDF prevents apoptotic-like death (ALD) in the presence of DNA damaging agents, probably by reducing recA mRNA levels in a non-endonuclease-mediated manner (PubMed:22412352). Other studies (in strains BW25113 and MC4100, the latter makes EDF) demonstrate MazF does not cause PCD but instead bacteriostasis and possibly a dormant state as well as persister cell generation (PubMed:24375411).9 Publications

Enzyme regulationi

Inhibited by Mg2+ (PubMed:15537630). Stimulated in vitro in a concentration-dependent fashion by EDF, which is able to overcome inhibition by cognate antitoxin MazE (PubMed:21419338). The TA module is antagonized by stress response kinase SrkA, but probably not by phosphorylation of MazF (PubMed:23416055).3 Publications

GO - Molecular functioni

  • DNA binding Source: UniProtKB-KW
  • endoribonuclease activity Source: UniProtKB
  • RNA binding Source: UniProtKB-KW

GO - Biological processi

  • defense response to virus Source: UniProtKB-KW
  • mRNA catabolic process Source: UniProtKB
  • negative regulation of cell growth Source: UniProtKB
  • quorum sensing Source: UniProtKB-KW
  • regulation of transcription, DNA-templated Source: UniProtKB-KW
  • regulation of translation Source: UniProtKB-KW
  • rRNA catabolic process Source: UniProtKB
  • transcription, DNA-templated Source: UniProtKB-KW
Complete GO annotation...

Keywords - Molecular functioni

Endonuclease, Hydrolase, Nuclease, Repressor, Toxin

Keywords - Biological processi

Antiviral defense, Quorum sensing, Stress response, Transcription, Transcription regulation, Translation regulation

Keywords - Ligandi

DNA-binding, RNA-binding

Enzyme and pathway databases

BioCyciEcoCyc:EG11249-MONOMER.
ECOL316407:JW2753-MONOMER.
MetaCyc:EG11249-MONOMER.

Names & Taxonomyi

Protein namesi
Recommended name:
Endoribonuclease MazF (EC:3.1.27.-1 Publication)
Alternative name(s):
Toxin MazF
mRNA interferase MazF1 Publication
Gene namesi
Name:mazF
Synonyms:chpA, chpAK
Ordered Locus Names:b2782, JW2753
OrganismiEscherichia coli (strain K12)
Taxonomic identifieri83333 [NCBI]
Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacteralesEnterobacteriaceaeEscherichia
Proteomesi
  • UP000000318 Componenti: Chromosome
  • UP000000625 Componenti: Chromosome

Organism-specific databases

EcoGeneiEG11249. mazF.

Pathology & Biotechi

Biotechnological usei

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 expressed bacterial cellular protein can be the target, which can be produced for up to 4 days. The system also works in eukaryotic cells.1 Publication

Disruption phenotypei

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 disruption culture is lethal (PubMed:15316771). Cells missing mazE-mazF show reduced biofilm formation, and survive antibiotic treatment in log phase better than wild-type cells (PubMed:11222603, PubMed:19707553). However lag phase cells disrupted only for mazF had a lower survival rate than wild-type cells (PubMed:24375411). Cells missing mazE-mazF survive hydroxyurea treatment better than wild-type; further disruption of relE-relB and tonB yields even better survival (PubMed:20005847). Cells missing mazE-mazF undergo an apoptotic-like death (ALD) upon DNA damage characterized by membrane depolarization and DNA fragmentation; further disruption of recA prevents membrane depolarization (PubMed:22412352). Unlike the single srkA disruption mutant, a triple srkA-mazE-mazF disruption mutant shows no hyperlethality in the presence of nalidixic acid or UV light, suggesting SrkA has a negative effect on MazF (PubMed:23416055).10 Publications

Mutagenesis

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Mutagenesisi17 – 28FDPTK…EQAGH → GGGGGGGGGGG: Changes loop 1 to poly-G; loss of endoribonuclease activity. 1 PublicationAdd BLAST12
Mutagenesisi17 – 28FDPTK…EQAGH → LGPPSGSQPAKR: Changes loop 1 to MazF6 M.tuberculosis sequence; loss of endoribonuclease activity. 1 PublicationAdd BLAST12
Mutagenesisi17 – 28FDPTK…EQAGH → PDDSRGPVPSYS: Changes loop 1 to MazF M.xanthus sequence; loss of endoribonuclease activity. 1 PublicationAdd BLAST12
Mutagenesisi24E → A: Greatly reduces toxicity, about 10-fold less RNA cleavage activity. Expression in the presence of wt MazF has a dominant-negative phenotype, causing cell death as it titrates out the MazE antitoxin; still activates operon transcription. 2 Publications1
Mutagenesisi28H → A: No changes in toxicity. 1 Publication1
Mutagenesisi53 – 61TQSKGYPFE → GGGGGGGG or GGGGGGGGGGG: Changes loop 2 to poly-G; reduces endoribonuclease activity, alters cleavage sites. 1 Publication9
Mutagenesisi53 – 61TQSKGYPFE → SNLHRASEPGN: Changes loop 2 to MazF M.xanthus sequence; reduces endoribonuclease activity, alters cleavage sites. 1 Publication9
Mutagenesisi53 – 61TQSKGYPFE → SNTALAAMPGN: Changes loop 2 to MazF6 M.tuberculosis sequence; reduces endoribonuclease activity, alters cleavage sites. 1 Publication9

Chemistry databases

ChEMBLiCHEMBL1795096.

PTM / Processingi

Molecule processing

Feature keyPosition(s)DescriptionActionsGraphical viewLength
ChainiPRO_00002018971 – 111Endoribonuclease MazFAdd BLAST111

Proteomic databases

PaxDbiP0AE70.
PRIDEiP0AE70.

Expressioni

Inductioni

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 (PubMed:12972253), but also to not occur after amino acid starvation and to be regulated by ppGpp (PubMed:8650219). Also induced in M9 minimal medium and by chloramphenicol treatment (PubMed:21944167). 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 (PubMed:2844820, PubMed:8650219). Operon induced by ectopic expression of toxin RelE; operon induction by amino acid starvation requires the relBEF operon (PubMed:23432955).7 Publications

Interactioni

Subunit structurei

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. Binds to EDF but not a mutated EDF (NNGNN) (PubMed:21419338).5 Publications

Protein-protein interaction databases

BioGridi4262300. 13 interactors.
IntActiP0AE70. 2 interactors.
STRINGi511145.b2782.

Chemistry databases

BindingDBiP0AE70.

Structurei

Secondary structure

1111
Legend: HelixTurnBeta strandPDB Structure known for this area
Show more details
Feature keyPosition(s)DescriptionActionsGraphical viewLength
Beta strandi11 – 15Combined sources5
Turni19 – 22Combined sources4
Beta strandi29 – 33Combined sources5
Helixi37 – 43Combined sources7
Beta strandi46 – 53Combined sources8
Beta strandi61 – 63Combined sources3
Beta strandi66 – 69Combined sources4
Beta strandi72 – 74Combined sources3
Helixi75 – 77Combined sources3
Beta strandi79 – 81Combined sources3
Helixi83 – 86Combined sources4
Beta strandi89 – 93Combined sources5
Helixi96 – 110Combined sources15

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
PDB entryMethodResolution (Å)ChainPositionsPDBsum
1UB4X-ray1.70A/B2-111[»]
3NFCX-ray2.00A/B/C/D/E/F1-111[»]
5CK9X-ray1.90A/B1-111[»]
5CKBX-ray2.80A/B1-111[»]
5CKDX-ray1.70A/B1-111[»]
5CKEX-ray2.31A/B1-111[»]
5CKFX-ray2.80A/B1-111[»]
5CKHX-ray2.45A/B1-111[»]
5CO7X-ray3.49A/B/C/D/E/F1-111[»]
5CQXX-ray1.63A/B1-111[»]
5CQYX-ray2.48A/B1-111[»]
5CR2X-ray2.90A/B/C1-111[»]
DisProtiDP00299.
ProteinModelPortaliP0AE70.
SMRiP0AE70.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiP0AE70.

Family & Domainsi

Region

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Regioni17 – 28Loop 1, participates in catalytic activity1 PublicationAdd BLAST12
Regioni53 – 61Loop 2, involved in substrate recognition1 Publication9

Domaini

Loop 1 (residues 17-28) effects catalytic activity while recognition of the ACA cleavage site is influenced by loop 2 (residues 53-61). Alterations of loop 2 generate new cleavage sites in addition to retaining the original cleavage site.1 Publication

Sequence similaritiesi

Belongs to the PemK/MazF family.Curated

Phylogenomic databases

eggNOGiENOG4105MZG. Bacteria.
COG2337. LUCA.
HOGENOMiHOG000290185.
InParanoidiP0AE70.
KOiK07171.
OMAiQAGREQS.
PhylomeDBiP0AE70.

Family and domain databases

Gene3Di2.30.30.110. 1 hit.
InterProiIPR003477. PemK-like.
IPR011067. Plasmid_toxin/cell-grow_inhib.
[Graphical view]
PfamiPF02452. PemK_toxin. 1 hit.
[Graphical view]
SUPFAMiSSF50118. SSF50118. 1 hit.

Sequencei

Sequence statusi: Complete.

P0AE70-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MVSRYVPDMG DLIWVDFDPT KGSEQAGHRP AVVLSPFMYN NKTGMCLCVP
60 70 80 90 100
CTTQSKGYPF EVVLSGQERD GVALADQVKS IAWRARGATK KGTVAPEELQ
110
LIKAKINVLI G
Length:111
Mass (Da):12,098
Last modified:December 6, 2005 - v1
Checksum:i1579C867DD6B96AC
GO

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
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.
PIRiB49339.
RefSeqiNP_417262.1. NC_000913.3.
WP_000254738.1. NZ_LN832404.1.

Genome annotation databases

EnsemblBacteriaiAAC75824; AAC75824; b2782.
BAE76856; BAE76856; BAE76856.
GeneIDi947252.
KEGGiecj:JW2753.
eco:b2782.
PATRICi32120980. VBIEscCol129921_2882.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
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.
PIRiB49339.
RefSeqiNP_417262.1. NC_000913.3.
WP_000254738.1. NZ_LN832404.1.

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
PDB entryMethodResolution (Å)ChainPositionsPDBsum
1UB4X-ray1.70A/B2-111[»]
3NFCX-ray2.00A/B/C/D/E/F1-111[»]
5CK9X-ray1.90A/B1-111[»]
5CKBX-ray2.80A/B1-111[»]
5CKDX-ray1.70A/B1-111[»]
5CKEX-ray2.31A/B1-111[»]
5CKFX-ray2.80A/B1-111[»]
5CKHX-ray2.45A/B1-111[»]
5CO7X-ray3.49A/B/C/D/E/F1-111[»]
5CQXX-ray1.63A/B1-111[»]
5CQYX-ray2.48A/B1-111[»]
5CR2X-ray2.90A/B/C1-111[»]
DisProtiDP00299.
ProteinModelPortaliP0AE70.
SMRiP0AE70.
ModBaseiSearch...
MobiDBiSearch...

Protein-protein interaction databases

BioGridi4262300. 13 interactors.
IntActiP0AE70. 2 interactors.
STRINGi511145.b2782.

Chemistry databases

BindingDBiP0AE70.
ChEMBLiCHEMBL1795096.

Proteomic databases

PaxDbiP0AE70.
PRIDEiP0AE70.

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaiAAC75824; AAC75824; b2782.
BAE76856; BAE76856; BAE76856.
GeneIDi947252.
KEGGiecj:JW2753.
eco:b2782.
PATRICi32120980. VBIEscCol129921_2882.

Organism-specific databases

EchoBASEiEB1229.
EcoGeneiEG11249. mazF.

Phylogenomic databases

eggNOGiENOG4105MZG. Bacteria.
COG2337. LUCA.
HOGENOMiHOG000290185.
InParanoidiP0AE70.
KOiK07171.
OMAiQAGREQS.
PhylomeDBiP0AE70.

Enzyme and pathway databases

BioCyciEcoCyc:EG11249-MONOMER.
ECOL316407:JW2753-MONOMER.
MetaCyc:EG11249-MONOMER.

Miscellaneous databases

EvolutionaryTraceiP0AE70.
PROiP0AE70.

Family and domain databases

Gene3Di2.30.30.110. 1 hit.
InterProiIPR003477. PemK-like.
IPR011067. Plasmid_toxin/cell-grow_inhib.
[Graphical view]
PfamiPF02452. PemK_toxin. 1 hit.
[Graphical view]
SUPFAMiSSF50118. SSF50118. 1 hit.
ProtoNetiSearch...

Entry informationi

Entry nameiMAZF_ECOLI
AccessioniPrimary (citable) accession number: P0AE70
Secondary accession number(s): P33645, Q2MA50
Entry historyi
Integrated into UniProtKB/Swiss-Prot: December 6, 2005
Last sequence update: December 6, 2005
Last modified: November 2, 2016
This is version 88 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

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.1 Publication

Keywords - Technical termi

3D-structure, Complete proteome, Reference proteome

Documents

  1. Escherichia coli
    Escherichia coli (strain K12): entries and cross-references to EcoGene
  2. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references
  3. SIMILARITY comments
    Index of protein domains and families

Similar proteinsi

Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.