T2F1_PLAOK
ID T2F1_PLAOK Reviewed; 579 AA.
AC P14870;
DT 01-APR-1990, integrated into UniProtKB/Swiss-Prot.
DT 23-FEB-2022, sequence version 2.
DT 03-AUG-2022, entry version 119.
DE RecName: Full=Type II restriction enzyme FokI {ECO:0000303|PubMed:12654995};
DE Short=R.FokI {ECO:0000303|PubMed:2684765, ECO:0000303|PubMed:2784436};
DE EC=3.1.21.4 {ECO:0000269|PubMed:9724744};
DE AltName: Full=Endonuclease FokI;
DE AltName: Full=Type-2 restriction enzyme FokI;
GN Name=fokIR {ECO:0000303|PubMed:2684765}; Synonyms=rfoKI;
OS Planomicrobium okeanokoites (Planococcus okeanokoites) (Flavobacterium
OS okeanokoites).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Planococcaceae; Planomicrobium.
OX NCBI_TaxID=244;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 33414 / DSM 15489 / NBRC 12536 / NCIMB 561 / CIP 105082 / LMG
RC 4030 / VKM B-1175;
RX PubMed=2784436; DOI=10.1016/s0021-9258(18)83613-4;
RA Kita K., Kotani H., Sugisaki H., Takanami M.;
RT "The fokI restriction-modification system. I. Organization and nucleotide
RT sequences of the restriction and modification genes.";
RL J. Biol. Chem. 264:5751-5756(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 2-26.
RX PubMed=2684765; DOI=10.1016/0378-1119(89)90284-9;
RA Looney M.C., Moran L.S., Jack W.E., Feehery G.R., Benner J.S., Slatko B.E.,
RA Wilson G.G.;
RT "Nucleotide sequence of the FokI restriction-modification system: separate
RT strand-specificity domains in the methyltransferase.";
RL Gene 80:193-208(1989).
RN [3]
RP SUBUNIT.
RX PubMed=9724744; DOI=10.1073/pnas.95.18.10570;
RA Bitinaite J., Wah D.A., Aggarwal A.K., Schildkraut I.;
RT "FokI dimerization is required for DNA cleavage.";
RL Proc. Natl. Acad. Sci. U.S.A. 95:10570-10575(1998).
RN [4]
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 [5]
RP BIOTECHNOLOGY.
RX PubMed=24792163; DOI=10.1093/nar/gku329;
RA de Lange O., Wolf C., Dietze J., Elsaesser J., Morbitzer R., Lahaye T.;
RT "Programmable DNA-binding proteins from Burkholderia provide a fresh
RT perspective on the TALE-like repeat domain.";
RL Nucleic Acids Res. 42:7436-7449(2014).
RN [6]
RP BIOTECHNOLOGY USES REVIEW.
RX PubMed=25057889; DOI=10.1042/bj20140295;
RA Wright D.A., Li T., Yang B., Spalding M.H.;
RT "TALEN-mediated genome editing: prospects and perspectives.";
RL Biochem. J. 462:15-24(2014).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS), SUBUNIT, DOMAIN, DNA-BINDING, AND
RP ACTIVE SITE.
RC STRAIN=ATCC 33414 / DSM 15489 / NBRC 12536 / NCIMB 561 / CIP 105082 / LMG
RC 4030 / VKM B-1175;
RX PubMed=9214510; DOI=10.1038/40446;
RA Wah D.A., Hirsch J.A., Dorner L.F., Schildkraut I., Aggarwal A.K.;
RT "Structure of the multimodular endonuclease FokI bound to DNA.";
RL Nature 388:97-100(1997).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS).
RC STRAIN=ATCC 33414 / DSM 15489 / NBRC 12536 / NCIMB 561 / CIP 105082 / LMG
RC 4030 / VKM B-1175;
RX PubMed=9724743; DOI=10.1073/pnas.95.18.10564;
RA Wah D.A., Bitinaite J., Schildkraut I., Aggarwal A.K.;
RT "Structure of FokI has implications for DNA cleavage.";
RL Proc. Natl. Acad. Sci. U.S.A. 95:10564-10569(1998).
CC -!- FUNCTION: An S subtype restriction enzyme that recognizes the
CC asymmetric double-stranded sequence 5'-GGATG-3' and cleaves
CC respectively 14 bases after G-1 (top strand) and 13 bases before C-1
CC (bottom strand). {ECO:0000269|PubMed:9724744,
CC ECO:0000303|PubMed:12654995}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Endonucleolytic cleavage of DNA to give specific double-
CC stranded fragments with terminal 5'-phosphates.; EC=3.1.21.4;
CC Evidence={ECO:0000269|PubMed:9724744};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC -!- SUBUNIT: Monomer, in which form it can cleave DNA. Homodimer when bound
CC to DNA. {ECO:0000269|PubMed:9214510, ECO:0000269|PubMed:9724744}.
CC -!- DOMAIN: Has an N-terminal DNA recognition domain (approximately
CC residues 1-292) joined to an endonuclease domain (residues 389-583) by
CC a flexible linker. {ECO:0000303|PubMed:9214510}.
CC -!- BIOTECHNOLOGY: Adding the endonuclease domain C-terminal to different
CC DNA recognition domains from other enzymes allows generation of novel
CC restriction enzymes that are used to genetically engineer eukaryotic
CC cells. {ECO:0000269|PubMed:24792163, ECO:0000303|PubMed:25057889}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAA24927.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
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DR EMBL; J04623; AAA24927.1; ALT_INIT; Genomic_DNA.
DR EMBL; M28828; AAA24934.1; -; Genomic_DNA.
DR PDB; 1FOK; X-ray; 2.80 A; A=4-579.
DR PDB; 2FOK; X-ray; 2.30 A; A/B=1-579.
DR PDBsum; 1FOK; -.
DR PDBsum; 2FOK; -.
DR AlphaFoldDB; P14870; -.
DR REBASE; 1056; FokI.
DR BRENDA; 3.1.21.4; 2296.
DR EvolutionaryTrace; P14870; -.
DR PRO; PR:P14870; -.
DR GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR GO; GO:0009036; F:type II site-specific deoxyribonuclease activity; IEA:UniProtKB-EC.
DR GO; GO:0009307; P:DNA restriction-modification system; IEA:UniProtKB-KW.
DR Gene3D; 1.10.10.10; -; 1.
DR Gene3D; 3.90.241.10; -; 1.
DR InterPro; IPR015334; FokI_cleavage_dom.
DR InterPro; IPR004234; FokI_D1.
DR InterPro; IPR004233; FokI_D2.
DR InterPro; IPR031655; FokI_D3.
DR InterPro; IPR044945; FokI_dom_1_2.
DR InterPro; IPR011335; Restrct_endonuc-II-like.
DR InterPro; IPR036388; WH-like_DNA-bd_sf.
DR InterPro; IPR036390; WH_DNA-bd_sf.
DR Pfam; PF09254; Endonuc-FokI_C; 1.
DR Pfam; PF02980; FokI_C; 1.
DR Pfam; PF02981; FokI_N; 1.
DR Pfam; PF16902; Type2_restr_D3; 1.
DR SUPFAM; SSF46785; SSF46785; 3.
DR SUPFAM; SSF52980; SSF52980; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; DNA-binding; Endonuclease;
KW Hydrolase; Magnesium; Nuclease; Restriction system.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:2684765"
FT CHAIN 2..579
FT /note="Type II restriction enzyme FokI"
FT /id="PRO_0000077308"
FT ACT_SITE 450
FT /evidence="ECO:0000303|PubMed:9214510"
FT ACT_SITE 467
FT /evidence="ECO:0000303|PubMed:9214510"
FT ACT_SITE 469
FT /evidence="ECO:0000303|PubMed:9214510"
FT HELIX 17..24
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 32..39
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 41..45
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 49..60
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 61..63
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 68..72
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 79..81
FT /evidence="ECO:0007829|PDB:1FOK"
FT HELIX 87..91
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 98..101
FT /evidence="ECO:0007829|PDB:1FOK"
FT HELIX 104..116
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 119..123
FT /evidence="ECO:0007829|PDB:2FOK"
FT TURN 124..127
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 128..131
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 133..140
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 146..157
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 160..169
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 177..181
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 184..186
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 197..205
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 209..218
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 222..236
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 239..242
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 245..248
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 258..267
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 269..279
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 282..284
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 293..295
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 303..319
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 321..324
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 325..334
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 341..352
FT /evidence="ECO:0007829|PDB:2FOK"
FT TURN 353..355
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 358..361
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 364..367
FT /evidence="ECO:0007829|PDB:2FOK"
FT TURN 377..380
FT /evidence="ECO:0007829|PDB:1FOK"
FT HELIX 382..384
FT /evidence="ECO:0007829|PDB:1FOK"
FT HELIX 389..398
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 402..404
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 406..410
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 411..414
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 418..420
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 421..434
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 439..442
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 445..448
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 450..454
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 456..460
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 463..469
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 479..494
FT /evidence="ECO:0007829|PDB:2FOK"
FT TURN 497..499
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 504..507
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 515..522
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 528..539
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 542..547
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 548..560
FT /evidence="ECO:0007829|PDB:2FOK"
FT HELIX 565..569
FT /evidence="ECO:0007829|PDB:2FOK"
FT STRAND 573..576
FT /evidence="ECO:0007829|PDB:2FOK"
SQ SEQUENCE 579 AA; 65741 MW; EA31FD789ACF32D6 CRC64;
MVSKIRTFGW VQNPGKFENL KRVVQVFDRN SKVHNEVKNI KIPTLVKESK IQKELVAIMN
QHDLIYTYKE LVGTGTSIRS EAPCDAIIQA TIADQGNKKG YIDNWSSDGF LRWAHALGFI
EYINKSDSFV ITDVGLAYSK SADGSAIEKE ILIEAISSYP PAIRILTLLE DGQHLTKFDL
GKNLGFSGES GFTSLPEGIL LDTLANAMPK DKGEIRNNWE GSSDKYARMI GGWLDKLGLV
KQGKKEFIIP TLGKPDNKEF ISHAFKITGE GLKVLRRAKG STKFTRVPKR VYWEMLATNL
TDKEYVRTRR ALILEILIKA GSLKIEQIQD NLKKLGFDEV IETIENDIKG LINTGIFIEI
KGRFYQLKDH ILQFVIPNRG VTKQLVKSEL EEKKSELRHK LKYVPHEYIE LIEIARNSTQ
DRILEMKVME FFMKVYGYRG KHLGGSRKPD GAIYTVGSPI DYGVIVDTKA YSGGYNLPIG
QADEMQRYVE ENQTRNKHIN PNEWWKVYPS SVTEFKFLFV SGHFKGNYKA QLTRLNHITN
CNGAVLSVEE LLIGGEMIKA GTLTLEEVRR KFNNGEINF
