ID MNMA_ECOLI Reviewed; 368 AA. AC P25745; P75964; DT 01-MAY-1992, integrated into UniProtKB/Swiss-Prot. DT 30-MAY-2000, sequence version 4. DT 04-NOV-2026, entry version 190. DE RecName: Full=tRNA-specific 2-thiouridylase MnmA; DE EC=2.8.1.13 {ECO:0000269|PubMed:12549933, ECO:0000269|PubMed:16387657}; GN Name=mnmA; Synonyms=asuE, trmU, ycfB; OrderedLocusNames=b1133, JW1119; OS Escherichia coli (strain K12). OC Bacteria; Pseudomonadati; Pseudomonadota; Gammaproteobacteria; OC Enterobacterales; Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911; RX PubMed=8905232; DOI=10.1093/dnares/3.3.137; RA Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K., RA Inada T., Itoh T., Kajihara M., Kanai K., Kashimoto K., Kimura S., RA Kitagawa M., Makino K., Masuda S., Miki T., Mizobuchi K., Mori H., RA Motomura K., Nakamura Y., Nashimoto H., Nishio Y., Saito N., Sampei G., RA Seki Y., Tagami H., Takemoto K., Wada C., Yamamoto Y., Yano M., RA Horiuchi T.; RT "A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to RT the 12.7-28.0 min region on the linkage map."; RL DNA Res. 3:137-155(1996). RN [2] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=9278503; DOI=10.1126/science.277.5331.1453; RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., RA Shao Y.; RT "The complete genome sequence of Escherichia coli K-12."; RL Science 277:1453-1462(1997). RN [3] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911; RX PubMed=16738553; DOI=10.1038/msb4100049; RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.; RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 RT and W3110."; RL Mol. Syst. Biol. 2:E1-E5(2006). RN [4] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 138-368. RC STRAIN=K12; RX PubMed=8969519; DOI=10.1099/13500872-142-11-3219; RA Green S.M., Malik T., Giles I.G., Drabble W.T.; RT "The purB gene of Escherichia coli K-12 is located in an operon."; RL Microbiology 142:3219-3230(1996). RN [5] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 254-368. RC STRAIN=K12; RX PubMed=1729205; DOI=10.1128/jb.174.1.130-136.1992; RA He B., Smith J.M., Zalkin H.; RT "Escherichia coli purB gene: cloning, nucleotide sequence, and regulation RT by purR."; RL J. Bacteriol. 174:130-136(1992). RN [6] RP MUTANT STUDIES. RC STRAIN=K12; RX PubMed=3881393; DOI=10.1128/jb.161.1.368-376.1985; RA Sullivan M.A., Cannon J.F., Webb F.H., Bock R.M.; RT "Antisuppressor mutation in Escherichia coli defective in biosynthesis of RT 5-methylaminomethyl-2-thiouridine."; RL J. Bacteriol. 161:368-376(1985). RN [7] RP FUNCTION, CATALYTIC ACTIVITY, ATP-BINDING, AND TRNA-BINDING. RC STRAIN=K12 / MC1061 / ATCC 53338 / DSM 7140; RX PubMed=12549933; DOI=10.1021/bi026536+; RA Kambampati R., Lauhon C.T.; RT "MnmA and IscS are required for in vitro 2-thiouridine biosynthesis in RT Escherichia coli."; RL Biochemistry 42:1109-1117(2003). RN [8] RP CATALYTIC ACTIVITY, AND INTERACTION WITH TUSE. RX PubMed=16387657; DOI=10.1016/j.molcel.2005.11.001; RA Ikeuchi Y., Shigi N., Kato J., Nishimura A., Suzuki T.; RT "Mechanistic insights into sulfur relay by multiple sulfur mediators RT involved in thiouridine biosynthesis at tRNA wobble positions."; RL Mol. Cell 21:97-108(2006). RN [9] RP CRYSTALLIZATION. RX PubMed=16582487; DOI=10.1107/s174430910600738x; RA Numata T., Ikeuchi Y., Fukai S., Adachi H., Matsumura H., Takano K., RA Murakami S., Inoue T., Mori Y., Sasaki T., Suzuki T., Nureki O.; RT "Crystallization and preliminary X-ray analysis of the tRNA thiolation RT enzyme MnmA from Escherichia coli complexed with tRNA(Glu)."; RL Acta Crystallogr. F 62:368-371(2006). RN [10] RP X-RAY CRYSTALLOGRAPHY IN COMPLEXES WITH AMP AND TRNA, ACTIVE SITE, RP DISULFIDE BOND, AND MUTAGENESIS OF ASP-17; MET-37; ASN-97; ASP-99; CYS-102; RP LYS-107; HIS-128; LYS-149; GLN-151; CYS-199; PHE-200; THR-239; ARG-311 AND RP GLN-344. RX PubMed=16871210; DOI=10.1038/nature04896; RA Numata T., Ikeuchi Y., Fukai S., Suzuki T., Nureki O.; RT "Snapshots of tRNA sulphuration via an adenylated intermediate."; RL Nature 442:419-424(2006). CC -!- FUNCTION: Catalyzes the 2-thiolation of uridine at the wobble position CC (U34) of tRNA(Lys), tRNA(Glu) and tRNA(Gln), leading to the formation CC of s(2)U34, the first step of tRNA-mnm(5)s(2)U34 synthesis. Sulfur is CC provided by IscS, via a sulfur-relay system. Binds ATP and its CC substrate tRNAs. {ECO:0000269|PubMed:12549933}. CC -!- CATALYTIC ACTIVITY: CC Reaction=S-sulfanyl-L-cysteinyl-[protein] + uridine(34) in tRNA + AH2 + CC ATP = 2-thiouridine(34) in tRNA + L-cysteinyl-[protein] + A + AMP + CC diphosphate + H(+); Xref=Rhea:RHEA:47032, Rhea:RHEA-COMP:10131, CC Rhea:RHEA-COMP:11726, Rhea:RHEA-COMP:11727, Rhea:RHEA-COMP:11728, CC ChEBI:CHEBI:13193, ChEBI:CHEBI:15378, ChEBI:CHEBI:17499, CC ChEBI:CHEBI:29950, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, CC ChEBI:CHEBI:61963, ChEBI:CHEBI:65315, ChEBI:CHEBI:87170, CC ChEBI:CHEBI:456215; EC=2.8.1.13; CC Evidence={ECO:0000269|PubMed:12549933, ECO:0000269|PubMed:16387657}; CC -!- SUBUNIT: Interacts with TusE. {ECO:0000269|PubMed:16387657}. CC -!- SUBCELLULAR LOCATION: Cytoplasm. CC -!- MISCELLANEOUS: During the reaction, ATP is used to activate the C2 atom CC of U34 by adenylation. After this, the persulfide sulfur on the CC catalytic cysteine is transferred to the C2 atom of U34. The reaction CC probably involves hydrogen sulfide that is generated from the CC persulfide intermediate and that acts as a nucleophile towards the CC activated C2 atom on U34. Subsequently, Cys-102 acts as a nucleophile CC towards Cys-199, and a transient disulfide bond is formed. CC -!- SIMILARITY: Belongs to the MnmA/TRMU family. {ECO:0000305}. CC -!- CAUTION: Was originally thought to be a 5-methylaminomethyl-2- CC methyltransferase involved in tRNA modification. {ECO:0000305}. CC -!- SEQUENCE CAUTION: CC Sequence=CAA41994.1; Type=Frameshift; Evidence={ECO:0000305}; CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; U00096; AAC74217.2; -; Genomic_DNA. DR EMBL; AP009048; BAA35955.1; -; Genomic_DNA. DR EMBL; X59307; CAA41994.1; ALT_FRAME; Genomic_DNA. DR EMBL; M74924; -; NOT_ANNOTATED_CDS; Genomic_DNA. DR PIR; B64858; B64858. DR RefSeq; NP_415651.4; NC_000913.3. DR RefSeq; WP_001297484.1; NZ_CP009789.1. DR PDB; 2DER; X-ray; 3.10 A; A/B=1-368. DR PDB; 2DET; X-ray; 3.40 A; A=1-368. DR PDB; 2DEU; X-ray; 3.40 A; A/B=1-368. DR AlphaFoldDB; P25745; -. DR MDposit; P25745; -. DR SMR; P25745; -. DR BioGRID; 4263129; 72. DR BioGRID; 850062; 1. DR ComplexPortal; CPX-2145; tRNA-specific 2-thiouridylase tusE-mnmA complex. DR DIP; DIP-11035N; -. DR FunCoup; P25745; 851. DR IntAct; P25745; 8. DR STRING; 511145.b1133; -. DR jPOST; P25745; -. DR PaxDb; 511145-b1133; -. DR EnsemblBacteria; AAC74217; AAC74217; b1133. DR GeneID; 75203719; -. DR GeneID; 945690; -. DR KEGG; ecj:JW1119; -. DR KEGG; eco:b1133; -. DR KEGG; ecoc:C3026_06815; -. DR PATRIC; fig|1411691.4.peg.1133; -. DR EchoBASE; EB1320; -. DR eggNOG; COG0482; Bacteria. DR HOGENOM; CLU_035188_1_0_6; -. DR InParanoid; P25745; -. DR OMA; AACHITT; -. DR OrthoDB; 9800696at2; -. DR PhylomeDB; P25745; -. DR BioCyc; EcoCyc:EG11344-MONOMER; -. DR BioCyc; MetaCyc:EG11344-MONOMER; -. DR BRENDA; 2.8.1.13; 2026. DR BRENDA; 2.8.1.4; 2026. DR BRENDA; 2.8.1.7; 2026. DR EvolutionaryTrace; P25745; -. DR PRO; PR:P25745; -. DR Proteomes; UP000000625; Chromosome. DR GO; GO:0005829; C:cytosol; IDA:EcoCyc. DR GO; GO:1990228; C:sulfurtransferase complex; IPI:ComplexPortal. DR GO; GO:0016783; F:sulfurtransferase activity; IDA:EcoCyc. DR GO; GO:0103016; F:tRNA-uridine 2-sulfurtransferase activity; IEA:UniProtKB-EC. DR GO; GO:0097532; P:stress response to acid chemical; IMP:EcoCyc. DR GO; GO:0002143; P:tRNA wobble position uridine thiolation; IDA:EcoCyc. DR CDD; cd01998; MnmA_TRMU-like; 1. DR FunFam; 2.30.30.280:FF:000001; tRNA-specific 2-thiouridylase MnmA; 1. DR FunFam; 2.40.30.10:FF:000023; tRNA-specific 2-thiouridylase MnmA; 1. DR FunFam; 3.40.50.620:FF:000004; tRNA-specific 2-thiouridylase MnmA; 1. DR Gene3D; 2.30.30.280; Adenine nucleotide alpha hydrolases-like domains; 1. DR Gene3D; 3.40.50.620; HUPs; 1. DR Gene3D; 2.40.30.10; Translation factors; 1. DR HAMAP; MF_00144; tRNA_thiouridyl_MnmA; 1. DR InterPro; IPR004506; MnmA-like. DR InterPro; IPR046885; MnmA-like_C. DR InterPro; IPR046884; MnmA-like_central. DR InterPro; IPR023382; MnmA-like_central_sf. DR InterPro; IPR014729; Rossmann-like_a/b/a_fold. DR NCBIfam; NF001138; mnmA; 1. DR NCBIfam; TIGR00420; trmU; 1. DR PANTHER; PTHR11933:SF5; MITOCHONDRIAL TRNA-SPECIFIC 2-THIOURIDYLASE 1; 1. DR PANTHER; PTHR11933; TRNA 5-METHYLAMINOMETHYL-2-THIOURIDYLATE -METHYLTRANSFERASE; 1. DR Pfam; PF03054; tRNA_Me_trans; 1. DR Pfam; PF20258; tRNA_Me_trans_C; 1. DR Pfam; PF20259; tRNA_Me_trans_M; 1. DR SUPFAM; SSF52402; Adenine nucleotide alpha hydrolases-like; 1. PE 1: Evidence at protein level; KW 3D-structure; ATP-binding; Cytoplasm; Disulfide bond; Nucleotide-binding; KW Reference proteome; RNA-binding; Transferase; tRNA processing; KW tRNA-binding. FT CHAIN 1..368 FT /note="tRNA-specific 2-thiouridylase MnmA" FT /id="PRO_0000121588" FT REGION 97..99 FT /note="Interaction with target base in tRNA" FT REGION 149..151 FT /note="Interaction with tRNA" FT REGION 243..252 FT /note="Interaction with tRNA" FT REGION 311..312 FT /note="Interaction with tRNA" FT ACT_SITE 102 FT /note="Nucleophile" FT /evidence="ECO:0000269|PubMed:16871210" FT ACT_SITE 199 FT /note="Cysteine persulfide intermediate" FT /evidence="ECO:0000269|PubMed:16871210" FT BINDING 11..18 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT BINDING 37 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT BINDING 127 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT SITE 128 FT /note="Interaction with tRNA" FT SITE 344 FT /note="Interaction with tRNA" FT DISULFID 102..199 FT /note="Alternate" FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 17 FT /note="D->A: Loss of activity." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 37 FT /note="M->A: Reduces activity by 60%." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 97 FT /note="N->A: Loss of activity." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 99 FT /note="D->A: Loss of activity." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 102 FT /note="C->A: Loss of activity." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 107 FT /note="K->M: Reduces activity by 75%." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 128 FT /note="H->A: Reduces activity by 90%." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 149 FT /note="K->A: Loss of activity." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 151 FT /note="Q->E: Loss of activity." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 199 FT /note="C->A: Abolishes the incorporation of sulfur from the FT sulfur-relay system; loss of activity." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 200 FT /note="F->A: Reduces activity by 60%." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 239 FT /note="T->A: Reduces activity by 50%." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 311 FT /note="R->A: Reduces activity by 75%." FT /evidence="ECO:0000269|PubMed:16871210" FT MUTAGEN 344 FT /note="Q->A: Loss of activity." FT /evidence="ECO:0000269|PubMed:16871210" FT STRAND 7..11 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 19..26 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 31..38 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 45..65 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 69..73 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 75..78 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 79..84 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 85..91 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 98..101 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 102..110 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 111..117 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 122..125 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 131..135 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 138..142 FT /evidence="ECO:0007829|PDB:2DER" FT TURN 147..149 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 152..155 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 160..165 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 170..172 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 175..184 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 198..204 FT /evidence="ECO:0007829|PDB:2DER" FT HELIX 206..211 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 220..223 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 224..226 FT /evidence="ECO:0007829|PDB:2DEU" FT STRAND 228..231 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 251..254 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 259..265 FT /evidence="ECO:0007829|PDB:2DER" FT TURN 266..269 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 270..276 FT /evidence="ECO:0007829|PDB:2DER" FT TURN 280..282 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 284..294 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 302..311 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 317..322 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 325..327 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 329..338 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 344..350 FT /evidence="ECO:0007829|PDB:2DER" FT STRAND 353..365 FT /evidence="ECO:0007829|PDB:2DER" SQ SEQUENCE 368 AA; 40959 MW; 22CA92676D0805CC CRC64; MSETAKKVIV GMSGGVDSSV SAWLLQQQGY QVEGLFMKNW EEDDGEEYCT AAADLADAQA VCDKLGIELH TVNFAAEYWD NVFELFLAEY KAGRTPNPDI LCNKEIKFKA FLEFAAEDLG ADYIATGHYV RRADVDGKSR LLRGLDSNKD QSYFLYTLSH EQIAQSLFPV GELEKPQVRK IAEDLGLVTA KKKDSTGICF IGERKFREFL GRYLPAQPGK IITVDGDEIG EHQGLMYHTL GQRKGLGIGG TKEGTEEPWY VVDKDVENNI LVVAQGHEHP RLMSVGLIAQ QLHWVDREPF TGTMRCTVKT RYRQTDIPCT VKALDDDRIE VIFDEPVAAV TPGQSAVFYN GEVCLGGGII EQRLPLPV //