ID T1MK_ECOLI Reviewed; 529 AA. AC P08957; Q2M5W7; DT 01-NOV-1988, integrated into UniProtKB/Swiss-Prot. DT 01-NOV-1988, sequence version 1. DT 27-MAR-2024, entry version 182. DE RecName: Full=Type I restriction enzyme EcoKI methylase subunit {ECO:0000305}; DE Short=M protein; DE EC=2.1.1.72 {ECO:0000269|PubMed:8514761}; DE AltName: Full=Type I methyltransferase M.EcoKI {ECO:0000303|PubMed:12654995}; DE Short=M.EcoKI {ECO:0000303|PubMed:12654995}; GN Name=hsdM {ECO:0000303|PubMed:6255295}; Synonyms=hsm; GN OrderedLocusNames=b4349, JW4312; OS Escherichia coli (strain K12). OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales; OC Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=K12; RX PubMed=3323532; DOI=10.1016/0022-2836(87)90303-2; RA Loenen W.A.M., Daniel A.S., Braymer H.D., Murray N.E.; RT "Organization and sequence of the hsd genes of Escherichia coli K-12."; RL J. Mol. Biol. 198:159-170(1987). RN [2] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=7610040; DOI=10.1093/nar/23.12.2105; RA Burland V.D., Plunkett G. III, Sofia H.J., Daniels D.L., Blattner F.R.; RT "Analysis of the Escherichia coli genome VI: DNA sequence of the region RT from 92.8 through 100 minutes."; RL Nucleic Acids Res. 23:2105-2119(1995). 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 1-5, FUNCTION, CATALYTIC ACTIVITY, AND SUBUNIT. RC STRAIN=K12; RX PubMed=8514761; DOI=10.1016/s0021-9258(19)38641-7; RA Dryden D.T., Cooper L.P., Murray N.E.; RT "Purification and characterization of the methyltransferase from the type 1 RT restriction and modification system of Escherichia coli K12."; RL J. Biol. Chem. 268:13228-13236(1993). RN [6] RP FUNCTION. RC STRAIN=K12; RX PubMed=4868368; DOI=10.1038/2171110a0; RA Meselson M., Yuan R.; RT "DNA restriction enzyme from E. coli."; RL Nature 217:1110-1114(1968). RN [7] RP FUNCTION, SUBUNIT, AND OPERON STRUCTURE. RC STRAIN=K12; RX PubMed=6255295; DOI=10.1007/bf00267350; RA Sain B., Murray N.E.; RT "The hsd (host specificity) genes of E. coli K 12."; RL Mol. Gen. Genet. 180:35-46(1980). RN [8] RP FUNCTION, AND SUBUNIT. RC STRAIN=K12; RX PubMed=9033396; DOI=10.1021/bi9619435; RA Dryden D.T., Cooper L.P., Thorpe P.H., Byron O.; RT "The in vitro assembly of the EcoKI type I DNA restriction/modification RT enzyme and its in vivo implications."; RL Biochemistry 36:1065-1076(1997). RN [9] RP IDENTIFICATION BY 2D-GEL. RX PubMed=9298644; DOI=10.1002/elps.1150180805; RA VanBogelen R.A., Abshire K.Z., Moldover B., Olson E.R., Neidhardt F.C.; RT "Escherichia coli proteome analysis using the gene-protein database."; RL Electrophoresis 18:1243-1251(1997). RN [10] RP INTERACTION WITH ESCHERICHIA PHAGE T7 PROTEIN OCR (MICROBIAL INFECTION). RC STRAIN=K12; RX PubMed=12235377; DOI=10.1093/nar/gkf518; RA Atanasiu C., Su T.J., Sturrock S.S., Dryden D.T.; RT "Interaction of the ocr gene 0.3 protein of bacteriophage T7 with EcoKI RT restriction/modification enzyme."; RL Nucleic Acids Res. 30:3936-3944(2002). RN [11] RP NOMENCLATURE, AND SUBTYPE. RX PubMed=12654995; DOI=10.1093/nar/gkg274; RA Roberts R.J., Belfort M., Bestor T., Bhagwat A.S., Bickle T.A., RA Bitinaite J., Blumenthal R.M., Degtyarev S.K., Dryden D.T., Dybvig K., RA Firman K., Gromova E.S., Gumport R.I., Halford S.E., Hattman S., RA Heitman J., Hornby D.P., Janulaitis A., Jeltsch A., Josephsen J., Kiss A., RA Klaenhammer T.R., Kobayashi I., Kong H., Krueger D.H., Lacks S., RA Marinus M.G., Miyahara M., Morgan R.D., Murray N.E., Nagaraja V., RA Piekarowicz A., Pingoud A., Raleigh E., Rao D.N., Reich N., Repin V.E., RA Selker E.U., Shaw P.C., Stein D.C., Stoddard B.L., Szybalski W., RA Trautner T.A., Van Etten J.L., Vitor J.M., Wilson G.G., Xu S.Y.; RT "A nomenclature for restriction enzymes, DNA methyltransferases, homing RT endonucleases and their genes."; RL Nucleic Acids Res. 31:1805-1812(2003). RN [12] {ECO:0007744|PDB:2AR0} RP X-RAY CRYSTALLOGRAPHY (2.80 ANGSTROMS) OF 2-527. RA Rajashankar K.R., Kniewel R., Lima C.D.; RT "Crystal structure of Type I restriction enzyme EcoKI M protein RT (EC 2.1.1.72) (M.EcoKI)."; RL Submitted (AUG-2005) to the PDB data bank. RN [13] {ECO:0007744|PDB:2Y7C, ECO:0007744|PDB:2Y7H} RP STRUCTURE BY ELECTRON MICROSCOPY (18.00 ANGSTROMS) IN COMPLEX WITH RP SPECIFICITY SUBUNIT AND ESCHERICHIA PHAGE T7 PROTEIN OCR, AND INTERACTION RP WITH ESCHERICHIA PHAGE T7 PROTEIN OCR (MICROBIAL INFECTION). RX PubMed=19074193; DOI=10.1093/nar/gkn988; RA Kennaway C.K., Obarska-Kosinska A., White J.H., Tuszynska I., Cooper L.P., RA Bujnicki J.M., Trinick J., Dryden D.T.; RT "The structure of M.EcoKI Type I DNA methyltransferase with a DNA mimic RT antirestriction protein."; RL Nucleic Acids Res. 37:762-770(2009). CC -!- FUNCTION: The subtype gamma methyltransferase (M) subunit of a type I CC restriction enzyme. The M and S subunits together form a CC methyltransferase (MTase) that methylates A-2 on the top and A-3 on the CC bottom strand of the sequence 5'-AACN(6)GTGC-3'. In the presence of the CC R subunit the complex can also act as an endonuclease, binding to the CC same target sequence but cutting the DNA some distance from this site. CC Whether the DNA is cut or modified depends on the methylation state of CC the target sequence. When the target site is unmodified, the DNA is CC cut. When the target site is hemimethylated, the complex acts as a CC maintenance MTase modifying the DNA so that both strands become CC methylated. After locating a non-methylated recognition site, the CC enzyme complex serves as a molecular motor that translocates DNA in an CC ATP-dependent manner until a collision occurs that triggers cleavage. CC {ECO:0000269|PubMed:4868368, ECO:0000269|PubMed:6255295, CC ECO:0000269|PubMed:8514761, ECO:0000269|PubMed:9033396}. CC -!- CATALYTIC ACTIVITY: CC Reaction=a 2'-deoxyadenosine in DNA + S-adenosyl-L-methionine = an CC N(6)-methyl-2'-deoxyadenosine in DNA + H(+) + S-adenosyl-L- CC homocysteine; Xref=Rhea:RHEA:15197, Rhea:RHEA-COMP:12418, Rhea:RHEA- CC COMP:12419, ChEBI:CHEBI:15378, ChEBI:CHEBI:57856, ChEBI:CHEBI:59789, CC ChEBI:CHEBI:90615, ChEBI:CHEBI:90616; EC=2.1.1.72; CC Evidence={ECO:0000269|PubMed:8514761}; CC -!- SUBUNIT: The type I restriction/modification system is composed of CC three polypeptides R, M and S (PubMed:6255295). The restriction enzyme CC has stoichiometry R(2)M(2)S(1) (PubMed:9033396). The methyltransferase CC is composed of M(2)S(1) (PubMed:8514761, PubMed:9033396, CC PubMed:19074193). {ECO:0000269|PubMed:19074193, CC ECO:0000269|PubMed:6255295, ECO:0000269|PubMed:8514761, CC ECO:0000269|PubMed:9033396}. CC -!- SUBUNIT: (Microbial infection) Interacts with Escherichia phage T7 CC protein Ocr; this interaction leads to the inhibition of the CC methyltransferase restriction enzyme M.EcoKI composed of M(2)S(1). CC {ECO:0000269|PubMed:12235377, ECO:0000269|PubMed:19074193}. CC -!- INTERACTION: CC P08957; P12295: ung; NbExp=3; IntAct=EBI-878571, EBI-559403; CC -!- INDUCTION: Encoded in the hsd locus, in the order hsdR-hsdM-hsdS. There CC is a promoter upstream of hsdR and another between hsdR and hsdM CC (PubMed:6255295). This probably allows expression of the methylase CC enzyme before the restriction-specific subunit (Probable). CC {ECO:0000269|PubMed:6255295, ECO:0000305|PubMed:6255295}. CC -!- MISCELLANEOUS: Type I restriction and modification enzymes are complex, CC multifunctional systems which require ATP, S-adenosyl methionine and CC Mg(2+) as cofactors and, in addition to their endonucleolytic and CC methylase activities, are potent DNA-dependent ATPases. CC -!- SIMILARITY: Belongs to the N(4)/N(6)-methyltransferase 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; X06545; CAA29792.1; -; Genomic_DNA. DR EMBL; U14003; AAA97246.1; -; Genomic_DNA. DR EMBL; U00096; AAC77305.1; -; Genomic_DNA. DR EMBL; AP009048; BAE78339.1; -; Genomic_DNA. DR PIR; B30375; XYECHM. DR RefSeq; NP_418769.1; NC_000913.3. DR RefSeq; WP_001063204.1; NZ_LN832404.1. DR PDB; 2AR0; X-ray; 2.80 A; A/B=2-527. DR PDB; 2Y7C; EM; 18.00 A; B/C=1-529. DR PDB; 2Y7H; EM; 18.00 A; B/C=1-529. DR PDBsum; 2AR0; -. DR PDBsum; 2Y7C; -. DR PDBsum; 2Y7H; -. DR AlphaFoldDB; P08957; -. DR SMR; P08957; -. DR BioGRID; 4262768; 110. DR ComplexPortal; CPX-5628; Type I restriction-modification EcoKI complex. DR DIP; DIP-9943N; -. DR IntAct; P08957; 15. DR STRING; 511145.b4349; -. DR REBASE; 13379; M.EcoW3110ORF4339P. DR REBASE; 152630; M1.Ret561ORF1035P. DR REBASE; 152641; M.Rsp1341ORF1035P. DR REBASE; 152653; M.Rsp113ORF1037P. DR REBASE; 152691; M.Rph931ORF1047P. DR REBASE; 152704; M.Rph831ORF1044P. DR REBASE; 152714; M.Rsp741ORF1035P. DR REBASE; 152736; M.Rsp871ORF1035P. DR REBASE; 156146; M.BamRD77ORF2498P. DR REBASE; 204159; M.Bli1441ORF2992P. DR REBASE; 204719; M.Bsu333ORF2986P. DR REBASE; 205029; M.Bve72ORF2738P. DR REBASE; 205122; M.Bve1413ORF3003P. DR REBASE; 3387; M.EcoKI. DR REBASE; 441884; M.EcoBL21FORF4361P. DR jPOST; P08957; -. DR PaxDb; 511145-b4349; -. DR EnsemblBacteria; AAC77305; AAC77305; b4349. DR GeneID; 948872; -. DR KEGG; ecj:JW4312; -. DR KEGG; eco:b4349; -. DR PATRIC; fig|1411691.4.peg.2337; -. DR EchoBASE; EB0453; -. DR eggNOG; COG0286; Bacteria. DR HOGENOM; CLU_018284_2_0_6; -. DR InParanoid; P08957; -. DR OMA; GTFGFMI; -. DR OrthoDB; 9784823at2; -. DR PhylomeDB; P08957; -. DR BioCyc; EcoCyc:EG10458-MONOMER; -. DR BioCyc; MetaCyc:EG10458-MONOMER; -. DR BRENDA; 3.1.21.3; 2165. DR EvolutionaryTrace; P08957; -. DR PRO; PR:P08957; -. DR Proteomes; UP000000318; Chromosome. DR Proteomes; UP000000625; Chromosome. DR GO; GO:0005829; C:cytosol; IDA:EcoCyc. DR GO; GO:0019812; C:type I site-specific deoxyribonuclease complex; IPI:ComplexPortal. DR GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW. DR GO; GO:0008170; F:N-methyltransferase activity; IMP:EcoliWiki. DR GO; GO:0009007; F:site-specific DNA-methyltransferase (adenine-specific) activity; IEA:UniProtKB-EC. DR GO; GO:0009307; P:DNA restriction-modification system; IDA:ComplexPortal. DR Gene3D; 1.20.1260.30; -; 1. DR Gene3D; 3.40.50.150; Vaccinia Virus protein VP39; 1. DR InterPro; IPR022749; D12N6_MeTrfase_N. DR InterPro; IPR003356; DNA_methylase_A-5. DR InterPro; IPR002052; DNA_methylase_N6_adenine_CS. DR InterPro; IPR029063; SAM-dependent_MTases_sf. DR InterPro; IPR038333; T1MK-like_N_sf. DR PANTHER; PTHR42933; SLR6095 PROTEIN; 1. DR PANTHER; PTHR42933:SF4; TYPE I RESTRICTION ENZYME ECOKI METHYLASE SUBUNIT; 1. DR Pfam; PF12161; HsdM_N; 1. DR Pfam; PF02384; N6_Mtase; 1. DR PRINTS; PR00507; N12N6MTFRASE. DR SUPFAM; SSF53335; S-adenosyl-L-methionine-dependent methyltransferases; 1. DR PROSITE; PS00092; N6_MTASE; 1. PE 1: Evidence at protein level; KW 3D-structure; Direct protein sequencing; DNA-binding; KW Host-virus interaction; Methyltransferase; Reference proteome; KW Restriction system; S-adenosyl-L-methionine; Transferase. FT CHAIN 1..529 FT /note="Type I restriction enzyme EcoKI methylase subunit" FT /id="PRO_0000088022" FT BINDING 148..153 FT /ligand="S-adenosyl-L-methionine" FT /ligand_id="ChEBI:CHEBI:59789" FT /evidence="ECO:0000250|UniProtKB:Q89Z59" FT BINDING 178..180 FT /ligand="S-adenosyl-L-methionine" FT /ligand_id="ChEBI:CHEBI:59789" FT /evidence="ECO:0000250|UniProtKB:Q89Z59" FT BINDING 216 FT /ligand="S-adenosyl-L-methionine" FT /ligand_id="ChEBI:CHEBI:59789" FT /evidence="ECO:0000250|UniProtKB:Q89Z59" FT HELIX 5..18 FT /evidence="ECO:0007829|PDB:2AR0" FT TURN 19..21 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 27..43 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 46..49 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 56..60 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 64..78 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 80..82 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 84..90 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 100..111 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 153..163 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 171..173 FT /evidence="ECO:0007829|PDB:2AR0" FT TURN 177..179 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 180..190 FT /evidence="ECO:0007829|PDB:2AR0" FT TURN 191..198 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 201..209 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 211..217 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 219..230 FT /evidence="ECO:0007829|PDB:2AR0" FT TURN 231..233 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 238..240 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 242..247 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 252..255 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 260..265 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 288..299 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 300..311 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 312..316 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 320..331 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 332..339 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 345..347 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 352..360 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 374..379 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 388..391 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 395..397 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 399..405 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 427..429 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 433..435 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 442..444 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 446..451 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 452..457 FT /evidence="ECO:0007829|PDB:2AR0" FT STRAND 459..461 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 482..505 FT /evidence="ECO:0007829|PDB:2AR0" FT HELIX 511..521 FT /evidence="ECO:0007829|PDB:2AR0" SQ SEQUENCE 529 AA; 59307 MW; D41C02203747C965 CRC64; MNNNDLVAKL WKLCDNLRDG GVSYQNYVNE LASLLFLKMC KETGQEAEYL PEGYRWDDLK SRIGQEQLQF YRKMLVHLGE DDKKLVQAVF HNVSTTITEP KQITALVSNM DSLDWYNGAH GKSRDDFGDM YEGLLQKNAN ETKSGAGQYF TPRPLIKTII HLLKPQPREV VQDPAAGTAG FLIEADRYVK SQTNDLDDLD GDTQDFQIHR AFIGLELVPG TRRLALMNCL LHDIEGNLDH GGAIRLGNTL GSDGENLPKA HIVATNPPFG SAAGTNITRT FVHPTSNKQL CFMQHIIETL HPGGRAAVVV PDNVLFEGGK GTDIRRDLMD KCHLHTILRL PTGIFYAQGV KTNVLFFTKG TVANPNQDKN CTDDVWVYDL RTNMPSFGKR TPFTDEHLQP FERVYGEDPH GLSPRTEGEW SFNAEETEVA DSEENKNTDQ HLATSRWRKF SREWIRTAKS DSLDISWLKD KDSIDADSLP EPDVLAAEAM GELVQALSEL DALMRELGAS DEADLQRQLL EEAFGGVKE //