SYI2_PSEFL
ID SYI2_PSEFL Reviewed; 1030 AA.
AC Q8L1B1; Q8GM72; Q8RL58;
DT 06-DEC-2005, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 3.
DT 03-AUG-2022, entry version 90.
DE RecName: Full=Isoleucine--tRNA ligase 2;
DE EC=6.1.1.5;
DE AltName: Full=Isoleucyl-tRNA synthetase 2;
DE Short=IleRS 2;
GN Name=ileS2; Synonyms=mupM;
OS Pseudomonas fluorescens.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=294;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 49323 / NCIMB 10586;
RX PubMed=12652905; DOI=10.1080/1042517021000033399;
RA Rangaswamy V., Hernandez-Guzman G., Shufran K.A., Bender C.L.;
RT "Analysis of rILERS, an isoleucyl-tRNA synthetase gene associated with
RT mupirocin production by Pseudomonas fluorescens NCIMB 10586.";
RL DNA Seq. 13:343-351(2002).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 49323 / NCIMB 10586;
RA El-Sayed A.K., Hothersall J., Cooper S.M., Stephens E., Thomas C.M.;
RT "Nucleotide sequence of a 75 kb region of the chromosome of Pseudomonas
RT fluorescens NCIMB 10586 required for biosynthesis of the polyketide
RT antibiotic mupirocin.";
RL Submitted (JAN-2002) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP SEQUENCE REVISION.
RA Haines A.S., Stephens E., Hothersall J., Thomas C.M.;
RL Submitted (FEB-2014) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-13, AND RESISTANCE
RP TO MUPIROCIN.
RC STRAIN=ATCC 49323 / NCIMB 10586;
RX PubMed=12672810; DOI=10.1074/jbc.m302633200;
RA Yanagisawa T., Kawakami M.;
RT "How does Pseudomonas fluorescens avoid suicide from its antibiotic
RT pseudomonic acid? Evidence for two evolutionarily distinct isoleucyl-tRNA
RT synthetases conferring self-defense.";
RL J. Biol. Chem. 278:25887-25894(2003).
CC -!- FUNCTION: Catalyzes the attachment of isoleucine to tRNA(Ile). As IleRS
CC can inadvertently accommodate and process structurally similar amino
CC acids such as valine, to avoid such errors it has two additional
CC distinct tRNA(Ile)-dependent editing activities. One activity is
CC designated as 'pretransfer' editing and involves the hydrolysis of
CC activated Val-AMP. The other activity is designated 'posttransfer'
CC editing and involves deacylation of mischarged Val-tRNA(Ile) (By
CC similarity). {ECO:0000250}.
CC -!- FUNCTION: Confers high-level resistance to the antibiotic mupirocin
CC (pseudomonic acid A), an Ile-analog produced by P.fluorescens NCIMB
CC 10586 itself that competitively inhibits activation by Ile-tRNA
CC synthetase, thus inhibiting protein biosynthesis.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + L-isoleucine + tRNA(Ile) = AMP + diphosphate + L-
CC isoleucyl-tRNA(Ile); Xref=Rhea:RHEA:11060, Rhea:RHEA-COMP:9666,
CC Rhea:RHEA-COMP:9695, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:58045, ChEBI:CHEBI:78442, ChEBI:CHEBI:78528,
CC ChEBI:CHEBI:456215; EC=6.1.1.5;
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105; Evidence={ECO:0000250};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=10 uM for isoleucine;
CC KM=330 uM for ATP;
CC -!- SUBUNIT: Monomer. {ECO:0000250}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250}.
CC -!- DOMAIN: IleRS has two distinct active sites: one for aminoacylation and
CC one for editing. The misactivated valine is translocated from the
CC active site to the editing site, which sterically excludes the
CC correctly activated isoleucine. The single editing site contains two
CC valyl binding pockets, one specific for each substrate (Val-AMP or Val-
CC tRNA(Ile)) (By similarity). {ECO:0000250}.
CC -!- MISCELLANEOUS: P.fluorescens NCIMB 10586 possesses two distinct IleRSs
CC (IleRS-R1 and IleRS-R2), each with a different level of sensitivity to
CC mupirocin. Purified IleRs-R2 shows no sensitivity to mupirocin even at
CC a concentration of 5 mM, 100'000 fold higher than the Ki value of
CC IleRS-R1.
CC -!- SIMILARITY: Belongs to the class-I aminoacyl-tRNA synthetase family.
CC IleS type 2 subfamily. {ECO:0000305}.
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DR EMBL; AY079084; AAL85500.1; -; Genomic_DNA.
DR EMBL; AF318063; AAM12927.2; -; Genomic_DNA.
DR EMBL; AB062785; BAC07171.1; -; Genomic_DNA.
DR AlphaFoldDB; Q8L1B1; -.
DR SMR; Q8L1B1; -.
DR PRIDE; Q8L1B1; -.
DR BRENDA; 6.1.1.5; 5121.
DR SABIO-RK; Q8L1B1; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0002161; F:aminoacyl-tRNA editing activity; IEA:InterPro.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-UniRule.
DR GO; GO:0004822; F:isoleucine-tRNA ligase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0000049; F:tRNA binding; IEA:InterPro.
DR GO; GO:0008270; F:zinc ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0006428; P:isoleucyl-tRNA aminoacylation; IEA:UniProtKB-UniRule.
DR GO; GO:0046677; P:response to antibiotic; IEA:UniProtKB-KW.
DR CDD; cd07961; Anticodon_Ia_Ile_ABEc; 1.
DR Gene3D; 3.40.50.620; -; 2.
DR HAMAP; MF_02003; Ile_tRNA_synth_type2; 1.
DR InterPro; IPR001412; aa-tRNA-synth_I_CS.
DR InterPro; IPR002300; aa-tRNA-synth_Ia.
DR InterPro; IPR033709; Anticodon_Ile_ABEc.
DR InterPro; IPR002301; Ile-tRNA-ligase.
DR InterPro; IPR023586; Ile-tRNA-ligase_type2.
DR InterPro; IPR013155; M/V/L/I-tRNA-synth_anticd-bd.
DR InterPro; IPR014729; Rossmann-like_a/b/a_fold.
DR InterPro; IPR009080; tRNAsynth_Ia_anticodon-bd.
DR InterPro; IPR009008; Val/Leu/Ile-tRNA-synth_edit.
DR PANTHER; PTHR42780; PTHR42780; 1.
DR Pfam; PF08264; Anticodon_1; 1.
DR Pfam; PF00133; tRNA-synt_1; 1.
DR PRINTS; PR00984; TRNASYNTHILE.
DR SUPFAM; SSF47323; SSF47323; 1.
DR SUPFAM; SSF50677; SSF50677; 1.
DR TIGRFAMs; TIGR00392; ileS; 1.
DR PROSITE; PS00178; AA_TRNA_LIGASE_I; 1.
PE 1: Evidence at protein level;
KW Aminoacyl-tRNA synthetase; Antibiotic resistance; ATP-binding; Cytoplasm;
KW Direct protein sequencing; Ligase; Metal-binding; Nucleotide-binding;
KW Protein biosynthesis; Zinc.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:12672810"
FT CHAIN 2..1030
FT /note="Isoleucine--tRNA ligase 2"
FT /id="PRO_0000098556"
FT MOTIF 48..58
FT /note="'HIGH' region"
FT MOTIF 589..593
FT /note="'KMSKS' region"
FT BINDING 592
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT CONFLICT 175
FT /note="G -> R (in Ref. 1; AAL85500)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 1030 AA; 117875 MW; 2D930693BE1DC935 CRC64;
MSTEGSGPVR FPAMEDAVLE RWEKEKTFEQ SISAREGKPV YVFYDGPPFA TGLPHYGHIL
TSYIKDVIPR YQTMLGKQVP RRWGWDCHGL PVEFEVEKAM GFKSKRDILE FGVEQFNDEC
RELVLKYADD WRGFVNRMGR WVDFDGAYKT MDNDYMESVL WGFKTLHDKG HVYEGGKIVP
YCVRCQTVLS NFEARLDDAF RPRRDMSAYV KFRQQDRPDT FFLAWTTTPW TLPANVALAV
AADENYVCIE HGEERLWLAE GCLGGLFDEP VILERCTGAE LAGLRYLPVV GEVIDASAHR
VVTADFVQMG DGSGIVHIAP AFGEDDALLG QQYELPAPNP VRDDGTFSDA VAQYAGQNIF
EATPRILADL KSSGLLFKQE QIEHNYPHCW RCDNPLIYRA VESWFIRASA LREQLVENNS
QVNWVPEHVK EGRFGDWIRN ARDWAVSRNR FWGAPIPVWR CDQCGTVEVM GSIAQIEARS
GRKVEDLHVP HIDEHRFACQ CCEGTMSRVT GVFDCWFESG AMPFASRHYP FENKQEFEQT
FPADFIVEYL AQTRGWFYTM MVISTGCFEQ NPFKNAMCHG VILAKDGRKM SKRLKNYPNP
MDLMQTHGSD ALRVALLASP VCKGEDIKFS EESVRDVVRR YHLLFWNCLQ FYKTFTEIDQ
FSPSGDLGQP LDNVLDHYLL HELAALESDI KMWMESLDFS KIYSRIEVFI NVLSTWYLRL
NKARIWRDGL DDDKRQCYEV LHYALSNFAR LLAPFMPFLA EAVYTELGYA DSVHLQDWPS
IDRQYLSYEL ADEMSSLRNL IASVRNVRET NGVSQKFPLR SIRVAGIEQA VLERYAQFLE
EELNVKQVQW AADADEWAQP VVVLIFSLLG KRLGPAMKAV TTAVKAGEYV IDEQGGLVAA
GQTIQPHEFE RRLTVRDTLN NVGIVENMVV WLDLDIDASL KREGAVRELN RRLQDLRKKA
KLGYTEKVDI AVLGGAYVDE ILVHHEDWLK SQLLVQSLLR SDLEAPLAVD EVELPEGDPV
RIQLRRSVLA
