KAE1_PYRAB
ID KAE1_PYRAB Reviewed; 324 AA.
AC Q9UXT7; G8ZKV4;
DT 02-NOV-2001, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-2000, sequence version 1.
DT 03-AUG-2022, entry version 137.
DE RecName: Full=tRNA N6-adenosine threonylcarbamoyltransferase {ECO:0000255|HAMAP-Rule:MF_01446};
DE EC=2.3.1.234 {ECO:0000255|HAMAP-Rule:MF_01446, ECO:0000269|PubMed:23258706};
DE AltName: Full=N6-L-threonylcarbamoyladenine synthase {ECO:0000255|HAMAP-Rule:MF_01446};
DE Short=t(6)A synthase {ECO:0000255|HAMAP-Rule:MF_01446};
DE AltName: Full=Pa-Kae1;
DE AltName: Full=t(6)A37 threonylcarbamoyladenosine biosynthesis protein Kae1 {ECO:0000255|HAMAP-Rule:MF_01446};
DE AltName: Full=tRNA threonylcarbamoyladenosine biosynthesis protein Kae1 {ECO:0000255|HAMAP-Rule:MF_01446};
GN Name=kae1 {ECO:0000255|HAMAP-Rule:MF_01446}; OrderedLocusNames=PYRAB17710;
GN ORFNames=PAB1159;
OS Pyrococcus abyssi (strain GE5 / Orsay).
OC Archaea; Euryarchaeota; Thermococci; Thermococcales; Thermococcaceae;
OC Pyrococcus.
OX NCBI_TaxID=272844;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=GE5 / Orsay;
RX PubMed=12622808; DOI=10.1046/j.1365-2958.2003.03381.x;
RA Cohen G.N., Barbe V., Flament D., Galperin M., Heilig R., Lecompte O.,
RA Poch O., Prieur D., Querellou J., Ripp R., Thierry J.-C., Van der Oost J.,
RA Weissenbach J., Zivanovic Y., Forterre P.;
RT "An integrated analysis of the genome of the hyperthermophilic archaeon
RT Pyrococcus abyssi.";
RL Mol. Microbiol. 47:1495-1512(2003).
RN [2]
RP GENOME REANNOTATION.
RC STRAIN=GE5 / Orsay;
RX PubMed=22057919; DOI=10.1007/s00284-011-0035-x;
RA Gao J., Wang J.;
RT "Re-annotation of two hyperthermophilic archaea Pyrococcus abyssi GE5 and
RT Pyrococcus furiosus DSM 3638.";
RL Curr. Microbiol. 64:118-129(2012).
RN [3]
RP FUNCTION, IRON-BINDING, AND DNA-BINDING.
RX PubMed=19143597; DOI=10.1042/bst0370029;
RA Hecker A., Graille M., Madec E., Gadelle D., Le Cam E., van Tilbergh H.,
RA Forterre P.;
RT "The universal Kae1 protein and the associated Bud32 kinase (PRPK), a
RT mysterious protein couple probably essential for genome maintenance in
RT Archaea and Eukarya.";
RL Biochem. Soc. Trans. 37:29-35(2009).
RN [4]
RP FUNCTION IN T(6)A TRNA MODIFICATION, CATALYTIC ACTIVITY, SUBUNIT, AND
RP TRNA- AND RNA-BINDING.
RC STRAIN=GE5 / Orsay;
RX PubMed=23258706; DOI=10.1093/nar/gks1287;
RA Perrochia L., Crozat E., Hecker A., Zhang W., Bareille J., Collinet B.,
RA van Tilbeurgh H., Forterre P., Basta T.;
RT "In vitro biosynthesis of a universal t6A tRNA modification in Archaea and
RT Eukarya.";
RL Nucleic Acids Res. 41:1953-1964(2013).
RN [5]
RP FUNCTION IN THE KEOPS COMPLEX, SUBUNIT, AND MUTAGENESIS OF HIS-107 AND
RP ASP-159.
RX PubMed=23945934; DOI=10.1093/nar/gkt720;
RA Perrochia L., Guetta D., Hecker A., Forterre P., Basta T.;
RT "Functional assignment of KEOPS/EKC complex subunits in the biosynthesis of
RT the universal t6A tRNA modification.";
RL Nucleic Acids Res. 41:9484-9499(2013).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) IN COMPLEX WITH ATP AND IRON,
RP FUNCTION, COFACTOR, SUBUNIT, IRON-BINDING, DNA-BINDING, AND MUTAGENESIS OF
RP TYR-127.
RC STRAIN=GE5 / Orsay;
RX PubMed=17766251; DOI=10.1093/nar/gkm554;
RA Hecker A., Leulliot N., Gadelle D., Graille M., Justome A., Dorlet P.,
RA Brochier C., Quevillon-Cheruel S., Le Cam E., van Tilbeurgh H.,
RA Forterre P.;
RT "An archaeal orthologue of the universal protein Kae1 is an iron
RT metalloprotein which exhibits atypical DNA-binding properties and apurinic-
RT endonuclease activity in vitro.";
RL Nucleic Acids Res. 35:6042-6051(2007).
CC -!- FUNCTION: Required for the formation of a threonylcarbamoyl group on
CC adenosine at position 37 (t(6)A37) in tRNAs that read codons beginning
CC with adenine. Is a component of the KEOPS complex that is probably
CC involved in the transfer of the threonylcarbamoyl moiety of
CC threonylcarbamoyl-AMP (TC-AMP) to the N6 group of A37. Kae1 likely
CC plays a direct catalytic role in this reaction, but requires other
CC protein(s) of the complex to fulfill this activity. In vitro, binds
CC tRNA, ssRNA, both single- and double-stranded DNA, and exhibits a low
CC ATPase activity. {ECO:0000255|HAMAP-Rule:MF_01446,
CC ECO:0000269|PubMed:17766251, ECO:0000269|PubMed:19143597,
CC ECO:0000269|PubMed:23258706, ECO:0000269|PubMed:23945934}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=adenosine(37) in tRNA + L-threonylcarbamoyladenylate = AMP +
CC H(+) + N(6)-L-threonylcarbamoyladenosine(37) in tRNA;
CC Xref=Rhea:RHEA:37059, Rhea:RHEA-COMP:10162, Rhea:RHEA-COMP:10163,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:73682, ChEBI:CHEBI:74411,
CC ChEBI:CHEBI:74418, ChEBI:CHEBI:456215; EC=2.3.1.234;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_01446,
CC ECO:0000269|PubMed:23258706};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_01446,
CC ECO:0000269|PubMed:17766251};
CC Note=Binds 1 Fe(2+) ion per subunit. {ECO:0000255|HAMAP-Rule:MF_01446,
CC ECO:0000269|PubMed:17766251};
CC -!- SUBUNIT: Monomer. Component of the KEOPS complex that consists of Kae1,
CC Bud32, Cgi121 and Pcc1; the whole complex dimerizes.
CC {ECO:0000255|HAMAP-Rule:MF_01446, ECO:0000269|PubMed:17766251,
CC ECO:0000269|PubMed:23258706, ECO:0000269|PubMed:23945934}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_01446}.
CC -!- MISCELLANEOUS: The ATP binding pocket identified in the crystal
CC structure would actually engage the substrate TC-AMP.
CC -!- SIMILARITY: Belongs to the KAE1 / TsaD family. {ECO:0000255|HAMAP-
CC Rule:MF_01446}.
CC -!- CAUTION: Was originally (PubMed:17766251) thought to have endonuclease
CC activity, but it could not be confirmed with orthologs purified from
CC M.jannaschii (PubMed:18951093) and S. cerevisiae (PubMed:21183954).
CC {ECO:0000305|PubMed:17766251}.
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DR EMBL; AJ248288; CAB50676.1; -; Genomic_DNA.
DR EMBL; HE613800; CCE71245.1; -; Genomic_DNA.
DR PIR; F75029; F75029.
DR RefSeq; WP_010868890.1; NC_000868.1.
DR PDB; 2IVN; X-ray; 1.65 A; A=1-324.
DR PDB; 2IVO; X-ray; 2.90 A; A/B/C/D=1-324.
DR PDB; 2IVP; X-ray; 2.50 A; A=1-324.
DR PDBsum; 2IVN; -.
DR PDBsum; 2IVO; -.
DR PDBsum; 2IVP; -.
DR AlphaFoldDB; Q9UXT7; -.
DR SMR; Q9UXT7; -.
DR STRING; 272844.PAB1159; -.
DR EnsemblBacteria; CAB50676; CAB50676; PAB1159.
DR GeneID; 1496074; -.
DR KEGG; pab:PAB1159; -.
DR PATRIC; fig|272844.11.peg.1890; -.
DR eggNOG; arCOG01183; Archaea.
DR HOGENOM; CLU_023208_2_2_2; -.
DR OMA; MRIMCEE; -.
DR OrthoDB; 34679at2157; -.
DR PhylomeDB; Q9UXT7; -.
DR BRENDA; 2.3.1.234; 5242.
DR EvolutionaryTrace; Q9UXT7; -.
DR Proteomes; UP000000810; Chromosome.
DR Proteomes; UP000009139; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0000408; C:EKC/KEOPS complex; IDA:UniProtKB.
DR GO; GO:0005506; F:iron ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0061711; F:N(6)-L-threonylcarbamoyladenine synthase activity; IEA:UniProtKB-EC.
DR GO; GO:0003727; F:single-stranded RNA binding; IDA:UniProtKB.
DR GO; GO:0000049; F:tRNA binding; IDA:UniProtKB.
DR GO; GO:0002949; P:tRNA threonylcarbamoyladenosine modification; IDA:UniProtKB.
DR HAMAP; MF_01446; Kae1; 1.
DR InterPro; IPR043129; ATPase_NBD.
DR InterPro; IPR000905; Gcp-like_dom.
DR InterPro; IPR017861; KAE1/TsaD.
DR InterPro; IPR022449; Kae1_arc.
DR InterPro; IPR034680; Kae1_archaea_euk.
DR InterPro; IPR017860; Peptidase_M22_CS.
DR PANTHER; PTHR11735; PTHR11735; 1.
DR PANTHER; PTHR11735:SF14; PTHR11735:SF14; 1.
DR Pfam; PF00814; TsaD; 1.
DR PRINTS; PR00789; OSIALOPTASE.
DR SUPFAM; SSF53067; SSF53067; 1.
DR TIGRFAMs; TIGR03722; arch_KAE1; 1.
DR TIGRFAMs; TIGR00329; gcp_kae1; 1.
DR PROSITE; PS01016; GLYCOPROTEASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acyltransferase; Cytoplasm; Iron; Metal-binding; Transferase;
KW tRNA processing.
FT CHAIN 1..324
FT /note="tRNA N6-adenosine threonylcarbamoyltransferase"
FT /id="PRO_0000096981"
FT BINDING 107
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01446,
FT ECO:0000269|PubMed:17766251"
FT BINDING 111
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01446,
FT ECO:0000269|PubMed:17766251"
FT BINDING 127..131
FT /ligand="substrate"
FT /evidence="ECO:0000305"
FT BINDING 127
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01446,
FT ECO:0000269|PubMed:17766251"
FT BINDING 159
FT /ligand="substrate"
FT /evidence="ECO:0000305"
FT BINDING 172
FT /ligand="substrate"
FT /evidence="ECO:0000305"
FT BINDING 176
FT /ligand="substrate"
FT /evidence="ECO:0000305"
FT BINDING 257
FT /ligand="substrate"
FT /evidence="ECO:0000305"
FT BINDING 285
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01446,
FT ECO:0000269|PubMed:17766251"
FT MUTAGEN 107
FT /note="H->A: Abolishes iron binding. Reduces the tRNA
FT modification activity of the KEOPS complex by 90%."
FT /evidence="ECO:0000269|PubMed:23945934"
FT MUTAGEN 127
FT /note="Y->F: Loss of iron, but no change in DNA-binding."
FT /evidence="ECO:0000269|PubMed:17766251"
FT MUTAGEN 159
FT /note="D->A: Completely impairs the tRNA modification
FT activity of the KEOPS complex. Does not impair ATPase
FT activity of the complex."
FT /evidence="ECO:0000269|PubMed:23945934"
FT STRAND 3..7
FT /evidence="ECO:0007829|PDB:2IVN"
FT STRAND 9..18
FT /evidence="ECO:0007829|PDB:2IVN"
FT STRAND 23..30
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 40..62
FT /evidence="ECO:0007829|PDB:2IVN"
FT TURN 66..68
FT /evidence="ECO:0007829|PDB:2IVN"
FT STRAND 71..79
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 81..97
FT /evidence="ECO:0007829|PDB:2IVN"
FT STRAND 102..106
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 107..113
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 114..118
FT /evidence="ECO:0007829|PDB:2IVN"
FT STRAND 124..128
FT /evidence="ECO:0007829|PDB:2IVN"
FT STRAND 133..139
FT /evidence="ECO:0007829|PDB:2IVN"
FT STRAND 142..152
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 154..165
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 171..180
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 200..212
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 217..243
FT /evidence="ECO:0007829|PDB:2IVN"
FT STRAND 246..252
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 253..256
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 258..271
FT /evidence="ECO:0007829|PDB:2IVN"
FT STRAND 274..276
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 280..283
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 287..299
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 306..309
FT /evidence="ECO:0007829|PDB:2IVN"
FT HELIX 317..319
FT /evidence="ECO:0007829|PDB:2IVN"
SQ SEQUENCE 324 AA; 35368 MW; D0A3E4279EA04C31 CRC64;
MLALGIEGTA HTLGIGIVSE DKVLANVFDT LTTEKGGIHP KEAAEHHARL MKPLLRKALS
EAGVSLDDID VIAFSQGPGL GPALRVVATA ARALAVKYRK PIVGVNHCIA HVEITKMFGV
KDPVGLYVSG GNTQVLALEG GRYRVFGETL DIGIGNAIDV FARELGLGFP GGPKVEKLAE
KGEKYIELPY AVKGMDLSFS GLLTEAIRKY RSGKYRVEDL AYSFQETAFA ALVEVTERAV
AHTEKDEVVL VGGVAANNRL REMLRIMTED RGIKFFVPPY DLCRDNGAMI AYTGLRMYKA
GISFRLEETI VKQKFRTDEV EIVW
