RUSA_ECOLI
ID RUSA_ECOLI Reviewed; 120 AA.
AC P0AG74; P40116; Q2MBN1;
DT 20-DEC-2005, integrated into UniProtKB/Swiss-Prot.
DT 20-DEC-2005, sequence version 1.
DT 03-AUG-2022, entry version 118.
DE RecName: Full=Crossover junction endodeoxyribonuclease RusA;
DE EC=3.1.21.10;
DE AltName: Full=Holliday junction nuclease RusA;
DE AltName: Full=Holliday junction resolvase;
GN Name=rusA; Synonyms=rus, ybcP; OrderedLocusNames=b0550, JW0538;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=K12;
RX PubMed=8648624; DOI=10.1006/jmbi.1996.0185;
RA Mahdi A.A., Sharples G.J., Mandal T.N., Lloyd R.G.;
RT "Holliday junction resolvases encoded by homologous rusA genes in
RT Escherichia coli K-12 and phage 82.";
RL J. Mol. Biol. 257:561-573(1996).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RA Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M.,
RA Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D.,
RA Namath A., Oefner P., Roberts D., Schramm S., Davis R.W.;
RT "Sequence of minutes 4-25 of Escherichia coli.";
RL Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases.
RN [3]
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 [4]
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 [5]
RP PROTEIN SEQUENCE OF 1-9, AND CHARACTERIZATION.
RX PubMed=7813450; DOI=10.1002/j.1460-2075.1994.tb06960.x;
RA Sharples G.J., Chan S.N., Mahdi A.A., Whitby M.C., Lloyd R.G.;
RT "Processing of intermediates in recombination and DNA repair:
RT identification of a new endonuclease that specifically cleaves Holliday
RT junctions.";
RL EMBO J. 13:6133-6142(1994).
RN [6]
RP CHARACTERIZATION, AND SUBUNIT.
RX PubMed=9169457; DOI=10.1074/jbc.272.23.14873;
RA Chan S.N., Harris L., Bolt E.L., Whitby M.C., Lloyd R.G.;
RT "Sequence specificity and biochemical characterization of the RusA Holliday
RT junction resolvase of Escherichia coli.";
RL J. Biol. Chem. 272:14873-14882(1997).
RN [7]
RP CLEAVAGE SPECIFICITY, DNA-BINDING, AND MUTAGENESIS OF ASP-70.
RX PubMed=9571038; DOI=10.1006/jmbi.1998.1681;
RA Giraud-Panis M.-J.E., Lilley D.M.J.;
RT "Structural recognition and distortion by the DNA junction-resolving enzyme
RT RusA.";
RL J. Mol. Biol. 278:117-133(1998).
RN [8]
RP MUTAGENESIS OF ASP-70; ASP-72; ASP-80; ASP-90; ASP-91; GLU-111 AND GLU-116.
RX PubMed=9973560; DOI=10.1006/jmbi.1998.2499;
RA Bolt E.L., Sharples G.J., Lloyd R.G.;
RT "Identification of three aspartic acid residues essential for catalysis by
RT the RusA holliday junction resolvase.";
RL J. Mol. Biol. 286:403-415(1999).
RN [9]
RP MUTAGENESIS OF ARG-16; TYR-17; ARG-19; ARG-58; ARG-68; ARG-69; ASN-73;
RP LYS-76; PHE-87 AND LYS-101.
RX PubMed=11080453; DOI=10.1006/jmbi.2000.4196;
RA Bolt E.L., Sharples G.J., Lloyd R.G.;
RT "Analysis of conserved basic residues associated with DNA binding (Arg69)
RT and catalysis (Lys76) by the RusA holliday junction resolvase.";
RL J. Mol. Biol. 304:165-176(2000).
RN [10]
RP X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS), AND SUBUNIT.
RX PubMed=14656440; DOI=10.1016/j.str.2003.11.004;
RA Rafferty J.B., Bolt E.L., Muranova T.A., Sedelnikova S.E., Leonard P.,
RA Pasquo A., Baker P.J., Rice D.W., Sharples G.J., Lloyd R.G.;
RT "The structure of Escherichia coli RusA endonuclease reveals a new Holliday
RT junction DNA binding fold.";
RL Structure 11:1557-1567(2003).
RN [11]
RP X-RAY CRYSTALLOGRAPHY (1.2 ANGSTROMS) OF MUTANT ASN-70 IN COMPLEX WITH DNA,
RP AND SUBUNIT.
RX PubMed=17028102; DOI=10.1093/nar/gkl447;
RA Macmaster R., Sedelnikova S., Baker P.J., Bolt E.L., Lloyd R.G.,
RA Rafferty J.B.;
RT "RusA Holliday junction resolvase: DNA complex structure -- insights into
RT selectivity and specificity.";
RL Nucleic Acids Res. 34:5577-5584(2006).
CC -!- FUNCTION: Endonuclease that resolves Holliday junction intermediates
CC made during homologous genetic recombination and DNA repair. Exhibits
CC sequence and structure-selective cleavage of four-way DNA junctions,
CC where it introduces symmetrical nicks in two strands of the same
CC polarity at the 5' side of CC dinucleotides. Corrects the defects in
CC genetic recombination and DNA repair associated with inactivation of
CC ruvAB or ruvC.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Endonucleolytic cleavage at a junction such as a reciprocal
CC single-stranded crossover between two homologous DNA duplexes
CC (Holliday junction).; EC=3.1.21.10;
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000305};
CC Note=Binds 1 Mg(2+) ion per subunit. {ECO:0000305};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:14656440,
CC ECO:0000269|PubMed:17028102, ECO:0000269|PubMed:9169457}.
CC -!- MISCELLANEOUS: Encoded by the cryptic lambdoid prophage DLP12. Normally
CC not expressed. Complete suppression of ruv mutations by RusA is
CC dependent on the activity of RecG.
CC -!- SIMILARITY: Belongs to the RusA family. {ECO:0000305}.
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DR EMBL; X92587; CAA63321.1; -; Genomic_DNA.
DR EMBL; U82598; AAB40746.1; -; Genomic_DNA.
DR EMBL; U00096; AAC73651.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE76325.1; -; Genomic_DNA.
DR PIR; S66590; S66590.
DR RefSeq; NP_415082.1; NC_000913.3.
DR RefSeq; WP_001099712.1; NZ_LN832404.1.
DR PDB; 1Q8R; X-ray; 1.90 A; A/B=1-120.
DR PDB; 2H8C; X-ray; 3.10 A; A/B/C/D=1-120.
DR PDB; 2H8E; X-ray; 1.20 A; A=1-120.
DR PDBsum; 1Q8R; -.
DR PDBsum; 2H8C; -.
DR PDBsum; 2H8E; -.
DR AlphaFoldDB; P0AG74; -.
DR SMR; P0AG74; -.
DR BioGRID; 4261529; 59.
DR BioGRID; 849560; 4.
DR DIP; DIP-16988N; -.
DR IntAct; P0AG74; 1.
DR STRING; 511145.b0550; -.
DR PaxDb; P0AG74; -.
DR PRIDE; P0AG74; -.
DR EnsemblBacteria; AAC73651; AAC73651; b0550.
DR EnsemblBacteria; BAE76325; BAE76325; BAE76325.
DR GeneID; 945174; -.
DR KEGG; ecj:JW0538; -.
DR KEGG; eco:b0550; -.
DR PATRIC; fig|1411691.4.peg.1727; -.
DR EchoBASE; EB4170; -.
DR eggNOG; COG4570; Bacteria.
DR HOGENOM; CLU_139466_0_2_6; -.
DR InParanoid; P0AG74; -.
DR OMA; YISDWGK; -.
DR PhylomeDB; P0AG74; -.
DR BioCyc; EcoCyc:G6306-MON; -.
DR BRENDA; 3.1.21.10; 2026.
DR EvolutionaryTrace; P0AG74; -.
DR PRO; PR:P0AG74; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0008821; F:crossover junction endodeoxyribonuclease activity; IDA:EcoCyc.
DR GO; GO:0000400; F:four-way junction DNA binding; IDA:EcoCyc.
DR GO; GO:0000287; F:magnesium ion binding; IEA:InterPro.
DR GO; GO:0006310; P:DNA recombination; IEA:UniProtKB-KW.
DR GO; GO:0006281; P:DNA repair; IEA:UniProtKB-KW.
DR Gene3D; 3.30.1330.70; -; 1.
DR InterPro; IPR016281; Endonuclease_RusA.
DR InterPro; IPR008822; Endonuclease_RusA-like.
DR InterPro; IPR036614; RusA-like_sf.
DR Pfam; PF05866; RusA; 1.
DR PIRSF; PIRSF001007; RusA; 1.
DR SUPFAM; SSF103084; SSF103084; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; DNA damage; DNA recombination;
KW DNA repair; Endonuclease; Hydrolase; Magnesium; Metal-binding; Nuclease;
KW Reference proteome.
FT CHAIN 1..120
FT /note="Crossover junction endodeoxyribonuclease RusA"
FT /id="PRO_0000192004"
FT REGION 13..16
FT /note="DNA-binding"
FT REGION 66..73
FT /note="DNA-binding"
FT BINDING 70
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000305"
FT BINDING 72
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000305"
FT BINDING 91
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000305"
FT MUTAGEN 16
FT /note="R->Q: No effect on ability to promote DNA repair."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 17
FT /note="Y->L: No effect on ability to promote DNA repair."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 19
FT /note="R->Q: No effect on ability to promote DNA repair."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 58
FT /note="R->Q: No effect on ability to promote DNA repair."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 68
FT /note="R->Q: No effect on ability to promote DNA repair."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 69
FT /note="R->Q,A: Loss of ability to promote DNA repair. Great
FT loss of Holliday junction resolvase activity. No effect on
FT DNA binding."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 70
FT /note="D->N: Reduces junction resolution 80-fold. No effect
FT on DNA binding."
FT /evidence="ECO:0000269|PubMed:9571038,
FT ECO:0000269|PubMed:9973560"
FT MUTAGEN 72
FT /note="D->N: Loss of ability to resolve junctions. No
FT effect on DNA binding."
FT /evidence="ECO:0000269|PubMed:9973560"
FT MUTAGEN 73
FT /note="N->A: Slight reduction in ability to promote DNA
FT repair. Great reduction in Holliday junction resolution
FT activity."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 76
FT /note="K->Q: Loss of ability to promote DNA repair. Loss of
FT Holliday junction resolvase activity. No effect on DNA
FT binding."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 76
FT /note="K->R: Loss of ability to promote DNA repair. Less
FT than 2% activity of Holliday junction resolvase. No effect
FT on DNA binding."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 80
FT /note="D->N: Slight reduction in ability to resolve
FT junctions. No effect on DNA binding."
FT /evidence="ECO:0000269|PubMed:9973560"
FT MUTAGEN 87
FT /note="F->Y,V: No effect on ability to promote DNA repair."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 90
FT /note="D->N: Slight reduction in ability to resolve
FT junctions. No effect on DNA binding."
FT /evidence="ECO:0000269|PubMed:9973560"
FT MUTAGEN 91
FT /note="D->N: Loss of ability to resolve junctions. No
FT effect on DNA binding."
FT /evidence="ECO:0000269|PubMed:9973560"
FT MUTAGEN 101
FT /note="K->Q: No effect on ability to promote DNA repair."
FT /evidence="ECO:0000269|PubMed:11080453"
FT MUTAGEN 111
FT /note="E->Q: No effect on resolvase activity or DNA
FT binding."
FT /evidence="ECO:0000269|PubMed:9973560"
FT MUTAGEN 116
FT /note="E->Q: No effect on resolvase activity or DNA
FT binding."
FT /evidence="ECO:0000269|PubMed:9973560"
FT STRAND 3..9
FT /evidence="ECO:0007829|PDB:2H8E"
FT HELIX 14..17
FT /evidence="ECO:0007829|PDB:2H8E"
FT STRAND 18..23
FT /evidence="ECO:0007829|PDB:2H8E"
FT STRAND 25..27
FT /evidence="ECO:0007829|PDB:2H8E"
FT HELIX 29..44
FT /evidence="ECO:0007829|PDB:2H8E"
FT STRAND 55..62
FT /evidence="ECO:0007829|PDB:2H8E"
FT STRAND 64..67
FT /evidence="ECO:0007829|PDB:2H8E"
FT HELIX 72..84
FT /evidence="ECO:0007829|PDB:2H8E"
FT STRAND 87..89
FT /evidence="ECO:0007829|PDB:2H8E"
FT HELIX 91..93
FT /evidence="ECO:0007829|PDB:2H8E"
FT STRAND 94..102
FT /evidence="ECO:0007829|PDB:2H8E"
FT STRAND 109..116
FT /evidence="ECO:0007829|PDB:2H8E"
SQ SEQUENCE 120 AA; 13846 MW; 38401669E8819EA6 CRC64;
MNTYSITLPW PPSNNRYYRH NRGRTHVSAE GQAYRDNVAR IIKNAMLDIG LAMPVKIRIE
CHMPDRRRRD LDNLQKAAFD ALTKAGFWLD DAQVVDYRVV KMPVTKGGRL ELTITEMGNE
