ID NUCA_SERMA Reviewed; 266 AA. AC P13717; DT 01-JAN-1990, integrated into UniProtKB/Swiss-Prot. DT 01-FEB-1996, sequence version 2. DT 03-MAY-2023, entry version 135. DE RecName: Full=Nuclease; DE EC=3.1.30.2; DE AltName: Full=Endonuclease; DE Contains: DE RecName: Full=Nuclease isoform Sm2; DE Contains: DE RecName: Full=Nuclease isoform Sm3; DE Contains: DE RecName: Full=Nuclease isoform Sm1; DE Flags: Precursor; GN Name=nucA; Synonyms=nuc; OS Serratia marcescens. OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales; OC Yersiniaceae; Serratia. OX NCBI_TaxID=615; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=W225; RX PubMed=3319779; DOI=10.1016/0378-1119(87)90121-1; RA Ball T.K., Saurugger P.N., Benedick M.J.; RT "The extracellular nuclease gene of Serratia marcescens and its secretion RT from Escherichia coli."; RL Gene 57:183-192(1987). RN [2] RP SEQUENCE REVISION TO 7-11. RA Benedick M.J.; RL Submitted (OCT-1989) to the EMBL/GenBank/DDBJ databases. RN [3] RP PARTIAL PROTEIN SEQUENCE, AND DISULFIDE BONDS. RX PubMed=2665765; DOI=10.1007/bf02910469; RA Biedermann K., Jepsen P.K., Riise E., Svendsen I.; RT "Purification and characterization of a Serratia marcescens nuclease RT produced by Escherichia coli."; RL Carlsberg Res. Commun. 54:17-27(1989). RN [4] RP IDENTIFICATION OF ISOFORMS SM1; SM2 AND SM3, MASS SPECTROMETRY, AND RP DISULFIDE BONDS. RC STRAIN=B10M1; RX PubMed=8373817; DOI=10.1016/0167-4838(93)90057-x; RA Pedersen J., Filimonova M.N., Roepstorff P., Biedermann K.; RT "Characterization of Serratia marcescens nuclease isoforms by plasma RT desorption mass spectrometry."; RL Biochim. Biophys. Acta 1202:13-21(1993). RN [5] RP ACTIVE SITE. RX PubMed=8078761; DOI=10.1093/nar/22.16.3280; RA Friedhoff P., Gimadutdinow O., Pingoud A.; RT "Identification of catalytically relevant amino acids of the extracellular RT Serratia marcescens endonuclease by alignment-guided mutagenesis."; RL Nucleic Acids Res. 22:3280-3287(1994). RN [6] RP BIOPHYSICOCHEMICAL PROPERTIES. RX PubMed=7804150; RA Filimonova M.N., Krause K.L., Benedik M.J.; RT "Kinetic studies of the Serratia marcescens extracellular nuclease RT isoforms."; RL Biochem. Mol. Biol. Int. 33:1229-1236(1994). RN [7] RP MUTAGENESIS OF ARG-78; ARG-108; HIS-110; ASN-140 AND GLU-148. RX PubMed=8758988; DOI=10.1093/nar/24.14.2632; RA Friedhoff P., Kolmes B., Gimadutdinow O., Wende W., Krause K.L., RA Pingoud A.; RT "Analysis of the mechanism of the Serratia nuclease using site-directed RT mutagenesis."; RL Nucleic Acids Res. 24:2632-2639(1996). RN [8] RP REVIEW. RX PubMed=9711834; DOI=10.1111/j.1574-6968.1998.tb13120.x; RA Benedik M.J., Strych U.; RT "Serratia marcescens and its extracellular nuclease."; RL FEMS Microbiol. Lett. 165:1-13(1998). RN [9] RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) IN COMPLEX WITH MAGNESIUM IONS, AND RP SUBUNIT. RX PubMed=7664065; DOI=10.1038/nsb0794-461; RA Miller M.D., Tanner J., Alpaugh M., Benedik M.J., Krause K.L.; RT "2.1-A structure of Serratia endonuclease suggests a mechanism for binding RT to double-stranded DNA."; RL Nat. Struct. Biol. 1:461-468(1994). RN [10] RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS). RX PubMed=9257723; DOI=10.1016/s0014-5793(97)00512-7; RA Lunin V.Y., Levdikov V.M., Shlyapnikov S.V., Blagova E.V., Lunin V.V., RA Wilson K.S., Mikhailov A.M.; RT "Three-dimensional structure of Serratia marcescens nuclease at 1.7-A RT resolution and mechanism of its action."; RL FEBS Lett. 412:217-222(1997). RN [11] RP X-RAY CRYSTALLOGRAPHY (1.1 ANGSTROMS) OF 22-266 IN COMPLEX WITH MAGNESIUM RP IONS. RX PubMed=10771425; DOI=10.1107/s090744490000322x; RA Shlyapnikov S.V., Lunin V.V., Perbandt M., Polyakov K.M., Lunin V.Y., RA Levdikov V.M., Betzel C., Mikhailov A.M.; RT "Atomic structure of the Serratia marcescens endonuclease at 1.1 A RT resolution and the enzyme reaction mechanism."; RL Acta Crystallogr. D 56:567-572(2000). CC -!- FUNCTION: Catalyzes the hydrolysis of both DNA and RNA, double- or CC single-stranded, at the 3'position of the phosphodiester bond to CC produce 5'-phosphorylated mono-, di-, tri- and tetranucleotides. DNA is CC a slightly better substrate than RNA. CC -!- CATALYTIC ACTIVITY: CC Reaction=Endonucleolytic cleavage to 5'-phosphomononucleotide and 5'- CC phosphooligonucleotide end-products.; EC=3.1.30.2; CC Evidence={ECO:0000255|PROSITE-ProRule:PRU10047}; CC -!- COFACTOR: CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; CC Note=Binds 1 Mg(2+) ion.; CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:10771425, CC ECO:0000269|PubMed:7664065}. CC -!- SUBCELLULAR LOCATION: Secreted. CC -!- MASS SPECTROMETRY: [Nuclease isoform Sm2]: Mass=26708.2; CC Method=Electrospray; Note=Isoform Sm2.; CC Evidence={ECO:0000269|PubMed:8373817}; CC -!- MASS SPECTROMETRY: [Nuclease isoform Sm3]: Mass=26591.8; CC Method=Electrospray; Note=Isoform Sm3.; CC Evidence={ECO:0000269|PubMed:8373817}; CC -!- MASS SPECTROMETRY: [Nuclease isoform Sm1]: Mass=26376.4; CC Method=Electrospray; Note=Isoform Sm1.; CC Evidence={ECO:0000269|PubMed:8373817}; CC -!- MISCELLANEOUS: The active site contains 1 hydrated magnesium ion that CC has only 1 direct interaction with the protein; all other interactions CC are via water molecules. CC -!- SIMILARITY: Belongs to the DNA/RNA non-specific endonuclease family. CC {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; M19495; AAA26560.1; -; Genomic_DNA. DR RefSeq; WP_015377376.1; NZ_WVHX01000034.1. DR PDB; 1G8T; X-ray; 1.10 A; A/B=22-266. DR PDB; 1QAE; X-ray; 2.05 A; A/B=22-266. DR PDB; 1QL0; X-ray; 1.10 A; A/B=26-266. DR PDB; 1SMN; X-ray; 2.04 A; A/B=22-266. DR PDB; 4E3Y; X-ray; 0.95 A; A/B=22-266. DR PDBsum; 1G8T; -. DR PDBsum; 1QAE; -. DR PDBsum; 1QL0; -. DR PDBsum; 1SMN; -. DR PDBsum; 4E3Y; -. DR AlphaFoldDB; P13717; -. DR SMR; P13717; -. DR STRING; 273526.SMDB11_1061; -. DR GeneID; 66715254; -. DR OrthoDB; 9811262at2; -. DR BRENDA; 3.1.30.2; 5690. DR EvolutionaryTrace; P13717; -. DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell. DR GO; GO:0004519; F:endonuclease activity; IEA:UniProtKB-KW. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0003676; F:nucleic acid binding; IEA:InterPro. DR CDD; cd00091; NUC; 1. DR Gene3D; 3.40.570.10; Extracellular Endonuclease, subunit A; 1. DR InterPro; IPR018524; DNA/RNA_endonuclease_AS. DR InterPro; IPR001604; DNA/RNA_non-sp_Endonuclease. DR InterPro; IPR044929; DNA/RNA_non-sp_Endonuclease_sf. DR InterPro; IPR020821; Extracellular_endonuc_su_A. DR InterPro; IPR044925; His-Me_finger_sf. DR InterPro; IPR040255; Non-specific_endonuclease. DR PANTHER; PTHR13966:SF5; ENDONUCLEASE G, MITOCHONDRIAL; 1. DR PANTHER; PTHR13966; ENDONUCLEASE RELATED; 1. DR Pfam; PF01223; Endonuclease_NS; 1. DR SMART; SM00892; Endonuclease_NS; 1. DR SMART; SM00477; NUC; 1. DR SUPFAM; SSF54060; His-Me finger endonucleases; 1. DR PROSITE; PS01070; NUCLEASE_NON_SPEC; 1. PE 1: Evidence at protein level; KW 3D-structure; Direct protein sequencing; Disulfide bond; Endonuclease; KW Hydrolase; Magnesium; Metal-binding; Nuclease; Secreted; Signal. FT SIGNAL 1..21 FT CHAIN 22..266 FT /note="Nuclease isoform Sm2" FT /id="PRO_0000019914" FT CHAIN 23..266 FT /note="Nuclease isoform Sm3" FT /id="PRO_0000019915" FT CHAIN 25..266 FT /note="Nuclease isoform Sm1" FT /id="PRO_0000019916" FT ACT_SITE 110 FT /note="Proton acceptor" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10047, FT ECO:0000269|PubMed:8078761" FT BINDING 140 FT /ligand="Mg(2+)" FT /ligand_id="ChEBI:CHEBI:18420" FT /ligand_note="catalytic" FT DISULFID 30..34 FT DISULFID 222..264 FT MUTAGEN 78 FT /note="R->A: Reduced activity 1000-fold." FT /evidence="ECO:0000269|PubMed:8758988" FT MUTAGEN 78 FT /note="R->K: Reduced activity 100-fold." FT /evidence="ECO:0000269|PubMed:8758988" FT MUTAGEN 108 FT /note="R->A: Reduced activity 1000-fold." FT /evidence="ECO:0000269|PubMed:8758988" FT MUTAGEN 108 FT /note="R->K: Reduced activity 50-fold." FT /evidence="ECO:0000269|PubMed:8758988" FT MUTAGEN 110 FT /note="H->A,D,E,K,Q: Reduced activity 100000-fold." FT /evidence="ECO:0000269|PubMed:8758988" FT MUTAGEN 110 FT /note="H->N: Reduced activity 1000-fold." FT /evidence="ECO:0000269|PubMed:8758988" FT MUTAGEN 140 FT /note="N->A,D,H: Reduced activity over 10000-fold." FT /evidence="ECO:0000269|PubMed:8758988" FT MUTAGEN 140 FT /note="N->Q: Reduced activity 1000-fold." FT /evidence="ECO:0000269|PubMed:8758988" FT MUTAGEN 148 FT /note="E->A,D,Q: Reduced activity over 800-fold." FT /evidence="ECO:0000269|PubMed:8758988" FT STRAND 29..32 FT /evidence="ECO:0007829|PDB:4E3Y" FT STRAND 39..42 FT /evidence="ECO:0007829|PDB:1QAE" FT STRAND 50..53 FT /evidence="ECO:0007829|PDB:4E3Y" FT TURN 55..57 FT /evidence="ECO:0007829|PDB:4E3Y" FT STRAND 58..67 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 88..90 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 94..97 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 100..104 FT /evidence="ECO:0007829|PDB:4E3Y" FT STRAND 106..112 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 114..117 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 123..127 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 129..131 FT /evidence="ECO:0007829|PDB:4E3Y" FT STRAND 132..136 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 137..140 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 143..153 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 154..156 FT /evidence="ECO:0007829|PDB:4E3Y" FT STRAND 163..170 FT /evidence="ECO:0007829|PDB:4E3Y" FT STRAND 189..197 FT /evidence="ECO:0007829|PDB:4E3Y" FT TURN 201..203 FT /evidence="ECO:0007829|PDB:4E3Y" FT STRAND 206..212 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 221..224 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 228..235 FT /evidence="ECO:0007829|PDB:4E3Y" FT STRAND 238..240 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 245..251 FT /evidence="ECO:0007829|PDB:4E3Y" FT HELIX 258..261 FT /evidence="ECO:0007829|PDB:4E3Y" SQ SEQUENCE 266 AA; 28945 MW; A0FF0C1430677B9E CRC64; MRFNNKMLAL AALLFAAQAS ADTLESIDNC AVGCPTGGSS NVSIVRHAYT LNNNSTTKFA NWVAYHITKD TPASGKTRNW KTDPALNPAD TLAPADYTGA NAALKVDRGH QAPLASLAGV SDWESLNYLS NITPQKSDLN QGAWARLEDQ ERKLIDRADI SSVYTVTGPL YERDMGKLPG TQKAHTIPSA YWKVIFINNS PAVNHYAAFL FDQNTPKGAD FCQFRVTVDE IEKRTGLIIW AGLPDDVQAS LKSKPGVLPE LMGCKN //