TNSA_ECOLX
ID TNSA_ECOLX Reviewed; 273 AA.
AC P13988; Q47662;
DT 01-JAN-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-JAN-1990, sequence version 1.
DT 03-AUG-2022, entry version 130.
DE RecName: Full=Transposon Tn7 transposition protein TnsA;
DE AltName: Full=Restriction enzyme-like endonuclease TnsA {ECO:0000305};
DE EC=3.1.21.- {ECO:0000305|PubMed:8947057};
GN Name=tnsA;
OS Escherichia coli.
OG Plasmid R721.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=562;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=2156235; DOI=10.1093/nar/18.4.901;
RA Flores C., Qadri M.I., Lichtenstein C.;
RT "DNA sequence analysis of five genes; tnsA, B, C, D and E, required for Tn7
RT transposition.";
RL Nucleic Acids Res. 18:901-911(1990).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=1655576; DOI=10.1016/0378-1119(91)90478-t;
RA Orle K.A., Craig N.L.;
RT "Identification of transposition proteins encoded by the bacterial
RT transposon Tn7.";
RL Gene 96:1-7(1990).
RN [3]
RP ERRATUM OF PUBMED:1655576.
RA Orle K.A., Craig N.L.;
RL Gene 104:125-131(1991).
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC PLASMID=R721;
RA Sampei G., Motomura K., Masuda S., Yamaguchi T., Ando K., Oishi T.,
RA Furuya N., Komano T., Mizobuchi K.;
RT "Organization and diversification of plasmid genomes: complete nucleotide
RT sequence of the R721 genome.";
RL Submitted (JUN-2000) to the EMBL/GenBank/DDBJ databases.
RN [5]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-134.
RX PubMed=3010949; DOI=10.1042/bj2340111;
RA Gay N.J., Tybulewicz V.L.J., Walker J.E.;
RT "Insertion of transposon Tn7 into the Escherichia coli glmS transcriptional
RT terminator.";
RL Biochem. J. 234:111-117(1986).
RN [6]
RP FUNCTION, MUTAGENESIS OF ASP-28; THR-34; GLU-37; HIS-57; SER-60; ASP-108;
RP ASP-114; GLU-144; GLU-147 AND GLU-149, COFACTOR, AND CATALYTIC ACTIVITY.
RX PubMed=8947057; DOI=10.1002/j.1460-2075.1996.tb01024.x;
RA Sarnovsky R.J., May E.W., Craig N.L.;
RT "The Tn7 transposase is a heteromeric complex in which DNA breakage and
RT joining activities are distributed between different gene products.";
RL EMBO J. 15:6348-6361(1996).
RN [7]
RP FUNCTION.
RX PubMed=10704304; DOI=10.1006/jmbi.2000.3558;
RA Biery M.C., Lopata M., Craig N.L.;
RT "A minimal system for Tn7 transposition: the transposon-encoded proteins
RT TnsA and TnsB can execute DNA breakage and joining reactions that generate
RT circularized Tn7 species.";
RL J. Mol. Biol. 297:25-37(2000).
RN [8]
RP SUBUNIT.
RX PubMed=23674682; DOI=10.1073/pnas.1305716110;
RA Choi K.Y., Li Y., Sarnovsky R., Craig N.L.;
RT "Direct interaction between the TnsA and TnsB subunits controls the
RT heteromeric Tn7 transposase.";
RL Proc. Natl. Acad. Sci. U.S.A. 110:E2038-E2045(2013).
RN [9] {ECO:0007744|PDB:1F1Z}
RP X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS) IN COMPLEX WITH MAGNESIUM, ACTIVE
RP SITE, AND COFACTOR.
RX PubMed=10911996; DOI=10.1016/s1097-2765(00)80267-1;
RA Hickman A.B., Li Y., Mathew S.V., May E.W., Craig N.L., Dyda F.;
RT "Unexpected structural diversity in DNA recombination: the restriction
RT endonuclease connection.";
RL Mol. Cell 5:1025-1034(2000).
RN [10] {ECO:0007744|PDB:1T0F}
RP X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) IN COMPLEX WITH MAGNESIUM, AND
RP INTERACTION WITH TNSC.
RX PubMed=15257292; DOI=10.1038/sj.emboj.7600311;
RA Ronning D.R., Li Y., Perez Z.N., Ross P.D., Hickman A.B., Craig N.L.,
RA Dyda F.;
RT "The carboxy-terminal portion of TnsC activates the Tn7 transposase through
RT a specific interaction with TnsA.";
RL EMBO J. 23:2972-2981(2004).
CC -!- FUNCTION: Required for Tn7 transposition. Forms the transposase,
CC together with TnsB (PubMed:8947057). TnsA executes the 5'-DNA strand
CC breakage reaction (PubMed:10704304). TnsABC and TnsD promote high-
CC frequency insertion of Tn7 into a specific target site known as ATT-Tn7
CC whereas TnsABC and TnsD promote low-frequency insertion into many
CC different sites. {ECO:0000269|PubMed:10704304,
CC ECO:0000269|PubMed:8947057}.
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:10911996, ECO:0000269|PubMed:15257292};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000305|PubMed:8947057};
CC Note=The active site binds 2 Mg(2+) ions (PubMed:10911996,
CC PubMed:15257292). According to PubMed:10704304, no transposition occurs
CC with Mg(2+) (PubMed:10704304). {ECO:0000269|PubMed:10704304,
CC ECO:0000269|PubMed:10911996, ECO:0000269|PubMed:15257292};
CC -!- SUBUNIT: Heteromer with TnsB (PubMed:23674682). Interacts with TnsC
CC (via C-terminus); this interaction allows TnsA to bind donor DNA
CC (PubMed:15257292). {ECO:0000269|PubMed:15257292,
CC ECO:0000269|PubMed:23674682}.
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; X17693; CAA35683.1; -; Genomic_DNA.
DR EMBL; M37391; AAA24680.1; -; Genomic_DNA.
DR EMBL; AP002527; BAB12613.1; -; Genomic_DNA.
DR EMBL; X03474; CAA27192.1; -; Genomic_DNA.
DR EMBL; V00620; CAA23895.1; -; Genomic_DNA.
DR PIR; S12637; S12637.
DR RefSeq; NP_065320.1; NC_002525.1.
DR RefSeq; WP_001029679.1; NZ_WWEV01000116.1.
DR PDB; 1F1Z; X-ray; 2.40 A; A/B=1-273.
DR PDB; 1T0F; X-ray; 1.85 A; A/B=1-273.
DR PDBsum; 1F1Z; -.
DR PDBsum; 1T0F; -.
DR AlphaFoldDB; P13988; -.
DR SMR; P13988; -.
DR IntAct; P13988; 1.
DR BRENDA; 2.7.7.B22; 2026.
DR EvolutionaryTrace; P13988; -.
DR GO; GO:0005694; C:chromosome; IDA:UniProtKB.
DR GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR GO; GO:0004519; F:endonuclease activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0004803; F:transposase activity; IDA:UniProtKB.
DR GO; GO:0006313; P:transposition, DNA-mediated; IDA:UniProtKB.
DR Gene3D; 1.10.10.10; -; 1.
DR Gene3D; 3.40.1350.10; -; 1.
DR InterPro; IPR011335; Restrct_endonuc-II-like.
DR InterPro; IPR014832; TnsA_C.
DR InterPro; IPR014833; TnsA_N.
DR InterPro; IPR011856; tRNA_endonuc-like_dom_sf.
DR InterPro; IPR036388; WH-like_DNA-bd_sf.
DR InterPro; IPR036390; WH_DNA-bd_sf.
DR Pfam; PF08721; Tn7_Tnp_TnsA_C; 1.
DR Pfam; PF08722; Tn7_Tnp_TnsA_N; 1.
DR SUPFAM; SSF46785; SSF46785; 1.
DR SUPFAM; SSF52980; SSF52980; 1.
PE 1: Evidence at protein level;
KW 3D-structure; DNA recombination; DNA-binding; Endonuclease; Hydrolase;
KW Magnesium; Manganese; Metal-binding; Nuclease; Plasmid;
KW Transposable element; Transposition.
FT CHAIN 1..273
FT /note="Transposon Tn7 transposition protein TnsA"
FT /id="PRO_0000072611"
FT DNA_BIND 90..108
FT /note="H-T-H motif"
FT /evidence="ECO:0000250"
FT ACT_SITE 63
FT /evidence="ECO:0000269|PubMed:10911996"
FT ACT_SITE 73
FT /evidence="ECO:0000250|UniProtKB:P15075"
FT ACT_SITE 114
FT /evidence="ECO:0000269|PubMed:10911996"
FT ACT_SITE 132
FT /evidence="ECO:0000269|PubMed:10911996"
FT BINDING 114
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:10911996,
FT ECO:0000269|PubMed:15257292"
FT BINDING 130
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:10911996,
FT ECO:0000269|PubMed:15257292"
FT BINDING 131
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:10911996,
FT ECO:0000269|PubMed:15257292"
FT MUTAGEN 28
FT /note="D->A: No effect on recombination."
FT /evidence="ECO:0000269|PubMed:8947057"
FT MUTAGEN 34
FT /note="T->A: No effect on recombination."
FT /evidence="ECO:0000269|PubMed:8947057"
FT MUTAGEN 37
FT /note="E->A: Modest decrease in 5'-cleavage."
FT /evidence="ECO:0000269|PubMed:8947057"
FT MUTAGEN 57
FT /note="H->A: Prevents recombination, may affect protein
FT structure."
FT /evidence="ECO:0000269|PubMed:8947057"
FT MUTAGEN 60
FT /note="S->A: Prevents recombination, may affect protein
FT structure."
FT /evidence="ECO:0000269|PubMed:8947057"
FT MUTAGEN 108
FT /note="D->A: No effect on recombination."
FT /evidence="ECO:0000269|PubMed:8947057"
FT MUTAGEN 114
FT /note="D->A: Prevents 5'-cleavage."
FT /evidence="ECO:0000269|PubMed:8947057"
FT MUTAGEN 144
FT /note="E->A: No effect on recombination."
FT /evidence="ECO:0000269|PubMed:8947057"
FT MUTAGEN 147
FT /note="E->A: No effect on recombination."
FT /evidence="ECO:0000269|PubMed:8947057"
FT MUTAGEN 149
FT /note="E->A: Decrease in 5' cleavage."
FT /evidence="ECO:0000269|PubMed:8947057"
FT CONFLICT 139
FT /note="D -> G (in Ref. 2; AAA24680)"
FT /evidence="ECO:0000305"
FT CONFLICT 227
FT /note="K -> Q (in Ref. 2; AAA24680)"
FT /evidence="ECO:0000305"
FT HELIX 10..18
FT /evidence="ECO:0007829|PDB:1T0F"
FT TURN 19..22
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 26..28
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 35..37
FT /evidence="ECO:0007829|PDB:1T0F"
FT STRAND 45..48
FT /evidence="ECO:0007829|PDB:1T0F"
FT TURN 50..52
FT /evidence="ECO:0007829|PDB:1T0F"
FT STRAND 53..58
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 61..72
FT /evidence="ECO:0007829|PDB:1T0F"
FT STRAND 76..84
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 87..97
FT /evidence="ECO:0007829|PDB:1T0F"
FT STRAND 112..123
FT /evidence="ECO:0007829|PDB:1T0F"
FT STRAND 125..131
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 134..138
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 140..156
FT /evidence="ECO:0007829|PDB:1T0F"
FT STRAND 160..163
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 165..167
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 170..180
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 189..192
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 195..204
FT /evidence="ECO:0007829|PDB:1T0F"
FT TURN 205..207
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 210..220
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 227..237
FT /evidence="ECO:0007829|PDB:1T0F"
FT STRAND 240..242
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 249..251
FT /evidence="ECO:0007829|PDB:1T0F"
FT HELIX 254..256
FT /evidence="ECO:0007829|PDB:1T0F"
FT STRAND 262..264
FT /evidence="ECO:0007829|PDB:1T0F"
SQ SEQUENCE 273 AA; 31257 MW; 2C2EFDE9A7D9C477 CRC64;
MAKANSSFSE VQIARRIKEG RGQGHGKDYI PWLTVQEVPS SGRSHRIYSH KTGRVHHLLS
DLELAVFLSL EWESSVLDIR EQFPLLPSDT RQIAIDSGIK HPVIRGVDQV MSTDFLVDCK
DGPFEQFAIQ VKPAAALQDE RTLEKLELER RYWQQKQIPW FIFTDKEINP VVKENIEWLY
SVKTEEVSAE LLAQLSPLAH ILQEKGDENI INVCKQVDIA YDLELGKTLS EIRALTANGF
IKFNIYKSFR ANKCADLCIS QVVNMEELRY VAN
