ID CAS2_ECOLI Reviewed; 94 AA. AC P45956; Q2MA75; DT 01-NOV-1995, integrated into UniProtKB/Swiss-Prot. DT 30-MAY-2000, sequence version 2. DT 29-OCT-2014, entry version 92. DE RecName: Full=CRISPR-associated endoribonuclease Cas2; DE EC=3.1.-.-; GN Name=ygbF; Synonyms=cas2; OrderedLocusNames=b2754, JW5438; OS Escherichia coli (strain K12). OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacteriales; OC Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=K12; RX PubMed=2656660; RA Nakata A., Amemura M., Makino K.; RT "Unusual nucleotide arrangement with repeated sequences in the RT Escherichia coli K-12 chromosome."; RL J. Bacteriol. 171:3553-3556(1989). RN [2] 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., RA Mau B., Shao Y.; RT "The complete genome sequence of Escherichia coli K-12."; RL Science 277:1453-1462(1997). RN [3] 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 RT MG1655 and W3110."; RL Mol. Syst. Biol. 2:E1-E5(2006). RN [4] RP IDENTIFICATION. RX PubMed=7567469; DOI=10.1093/nar/23.17.3554; RA Borodovsky M., McIninch J., Koonin E.V., Rudd K.E., Medigue C., RA Danchin A.; RT "Detection of new genes in a bacterial genome using Markov models for RT three gene classes."; RL Nucleic Acids Res. 23:3554-3562(1995). RN [5] RP OPERON STRUCTURE, AND INDUCTION BY LEUO. RC STRAIN=K12 / BW25113; RX PubMed=19429622; DOI=10.1128/JB.00108-09; RA Shimada T., Yamamoto K., Ishihama A.; RT "Involvement of the leucine response transcription factor LeuO in RT regulation of the genes for sulfa drug efflux."; RL J. Bacteriol. 191:4562-4571(2009). RN [6] RP INDUCTION BY H-NS. RC STRAIN=K12; RX PubMed=20132443; DOI=10.1111/j.1365-2958.2010.07073.x; RA Pul U., Wurm R., Arslan Z., Geissen R., Hofmann N., Wagner R.; RT "Identification and characterization of E. coli CRISPR-cas promoters RT and their silencing by H-NS."; RL Mol. Microbiol. 75:1495-1512(2010). RN [7] RP INDUCTION BY BAER, ROLE IN PLASMID SILENCING, AND DISRUPTION RP PHENOTYPE. RC STRAIN=K12 / BW25113; RX PubMed=21255106; DOI=10.1111/j.1365-2958.2010.07482.x; RA Perez-Rodriguez R., Haitjema C., Huang Q., Nam K.H., Bernardis S., RA Ke A., DeLisa M.P.; RT "Envelope stress is a trigger of CRISPR RNA-mediated DNA silencing in RT Escherichia coli."; RL Mol. Microbiol. 79:584-599(2011). RN [8] RP X-RAY CRYSTALLOGRAPHY (1.1 ANGSTROMS). RA Nocek B., Skarina T., Brown G., Yakunin A., Joachimiak A.; RT "Crystal structure of a putative ssRNA endonuclease Cas2, CRISPR RT adaptation protein from E.coli."; RL Submitted (AUG-2013) to the PDB data bank. RN [9] RP X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS) IN COMPLEX WITH CAS1, FUNCTION RP IN SPACER ACQUISITION, INTERACTION WITH CAS1 (YGBT), SUBUNIT, AND RP MUTAGENESIS OF GLU-9; ASN-10; ARG-14; ARG-16; ARG-18; ARG-27; GLU-65; RP 79-THR--VAL-94 AND ASP-84. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=24793649; DOI=10.1038/nsmb.2820; RA Nunez J.K., Kranzusch P.J., Noeske J., Wright A.V., Davies C.W., RA Doudna J.A.; RT "Cas1-Cas2 complex formation mediates spacer acquisition during RT CRISPR-Cas adaptive immunity."; RL Nat. Struct. Mol. Biol. 21:528-534(2014). CC -!- FUNCTION: CRISPR (clustered regularly interspaced short CC palindromic repeat), is an adaptive immune system that provides CC protection against mobile genetic elements (viruses, transposable CC elements and conjugative plasmids). CRISPR clusters contain CC sequences complementary to antecedent mobile elements and target CC invading nucleic acids. CRISPR clusters are transcribed and CC processed into CRISPR RNA (crRNA). The Cas1-Cas2 complex is CC involved in CRISPR adaptation, the first stage of CRISPR immunity, CC being required for the addition/removal of CRISPR spacers. CC Expression of Cas1-Cas2 in a strain lacking both genes permits CC spacer acquisition. This subunit's nuclease activity is not CC required for spacer acquisition. Does not seem to bind DNA alone; CC the Cas1-Cas2 complex preferentially binds CRISPR-locus DNA CC (PubMed:24793649). {ECO:0000269|PubMed:21255106, CC ECO:0000269|PubMed:24793649}. CC -!- SUBUNIT: Homodimer. Forms a Cas1-Cas2 heterotetramer, although a CC hexamer (2 Cas1 dimers and 1 Cas2 dimer) was the form crystallized CC and might be functional. {ECO:0000269|PubMed:24793649}. CC -!- INDUCTION: Repressed by H-NS, activated by LeuO. Activated by the CC BaeSR two-component regulatory system, possibly due to envelope CC stress. Part of the casABCDE-ygbT-ygbF operon. CC {ECO:0000269|PubMed:19429622, ECO:0000269|PubMed:20132443, CC ECO:0000269|PubMed:21255106}. CC -!- DISRUPTION PHENOTYPE: Loss of plasmid silencing. CC {ECO:0000269|PubMed:21255106}. CC -!- SIMILARITY: Belongs to the CRISPR-associated endoribonuclease Cas2 CC protein family. E.coli-subtype subfamily. {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; M27059; -; NOT_ANNOTATED_CDS; Unassigned_DNA. DR EMBL; U29579; AAA69264.1; -; Genomic_DNA. DR EMBL; U00096; AAC75796.2; -; Genomic_DNA. DR EMBL; AP009048; BAE76831.1; -; Genomic_DNA. DR PIR; F65056; F65056. DR RefSeq; NP_417234.2; NC_000913.3. DR RefSeq; YP_490963.1; NC_007779.1. DR PDB; 4MAK; X-ray; 1.10 A; A/B=1-94. DR PDB; 4P6I; X-ray; 2.30 A; A/B=1-94. DR PDB; 4QDL; X-ray; 2.70 A; E/F=1-94. DR PDBsum; 4MAK; -. DR PDBsum; 4P6I; -. DR PDBsum; 4QDL; -. DR ProteinModelPortal; P45956; -. DR SMR; P45956; 1-94. DR DIP; DIP-12109N; -. DR IntAct; P45956; 2. DR STRING; 511145.b2754; -. DR EnsemblBacteria; AAC75796; AAC75796; b2754. DR EnsemblBacteria; BAE76831; BAE76831; BAE76831. DR GeneID; 12930591; -. DR GeneID; 947213; -. DR KEGG; ecj:Y75_p2692; -. DR KEGG; eco:b2754; -. DR PATRIC; 32120918; VBIEscCol129921_2851. DR EchoBASE; EB2694; -. DR EcoGene; EG12845; ygbF. DR eggNOG; NOG12609; -. DR HOGENOM; HOG000015873; -. DR InParanoid; P45956; -. DR OMA; TNNEQGF; -. DR OrthoDB; EOG6ZWJJS; -. DR BioCyc; EcoCyc:EG12845-MONOMER; -. DR BioCyc; ECOL316407:JW5438-MONOMER; -. DR PRO; PR:P45956; -. DR Genevestigator; P45956; -. DR GO; GO:0004519; F:endonuclease activity; IEA:UniProtKB-KW. DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-KW. DR GO; GO:0043571; P:maintenance of CRISPR repeat elements; IMP:EcoCyc. DR InterPro; IPR010152; CRISPR-assoc_prot_Cas2_sub. DR Pfam; PF09707; Cas_Cas2CT1978; 1. DR TIGRFAMs; TIGR01873; cas_CT1978; 1. PE 1: Evidence at protein level; KW 3D-structure; Antiviral defense; Complete proteome; Endonuclease; KW Hydrolase; Nuclease; Reference proteome. FT CHAIN 1 94 CRISPR-associated endoribonuclease Cas2. FT /FTId=PRO_0000169312. FT MUTAGEN 9 9 E->A,R: No effect on spacer acquisition, FT Cas1-Cas2 complex formation or CRISPR FT DNA-binding by complex. FT {ECO:0000269|PubMed:24793649}. FT MUTAGEN 10 10 N->A: No effect on spacer acquisition. FT {ECO:0000269|PubMed:24793649}. FT MUTAGEN 14 14 R->A: Slight decrease in spacer FT acquisition. FT {ECO:0000269|PubMed:24793649}. FT MUTAGEN 16 16 R->A: Slight decrease in spacer FT acquisition. FT {ECO:0000269|PubMed:24793649}. FT MUTAGEN 18 18 R->A: Very little spacer acquisition. FT {ECO:0000269|PubMed:24793649}. FT MUTAGEN 27 27 R->A: Slight decrease in spacer FT acquisition. FT {ECO:0000269|PubMed:24793649}. FT MUTAGEN 65 65 E->A: No effect on spacer acquisition. FT {ECO:0000269|PubMed:24793649}. FT MUTAGEN 65 65 E->R: Slight decrease in spacer FT acquisition, Cas1-Cas2 complex formation FT or CRISPR DNA-binding by complex. Loss of FT spacer acquisition; when associated with FT R-84. {ECO:0000269|PubMed:24793649}. FT MUTAGEN 79 94 Missing: Loss of spacer acquisition, no FT Cas1-Cas2 complex formation, loss of FT CRISPR DNA-binding by complex. FT {ECO:0000269|PubMed:24793649}. FT MUTAGEN 84 84 D->A: No effect on spacer acquisition. FT {ECO:0000269|PubMed:24793649}. FT MUTAGEN 84 84 D->R: Slight decrease in spacer FT acquisition. Loss of spacer acquisition; FT when associated with R-65. FT {ECO:0000269|PubMed:24793649}. FT STRAND 3 10 {ECO:0000244|PDB:4MAK}. FT HELIX 13 22 {ECO:0000244|PDB:4MAK}. FT STRAND 23 27 {ECO:0000244|PDB:4MAK}. FT STRAND 30 34 {ECO:0000244|PDB:4MAK}. FT HELIX 37 50 {ECO:0000244|PDB:4MAK}. FT STRAND 55 61 {ECO:0000244|PDB:4MAK}. FT STRAND 63 66 {ECO:0000244|PDB:4MAK}. FT STRAND 68 73 {ECO:0000244|PDB:4MAK}. FT STRAND 79 83 {ECO:0000244|PDB:4P6I}. FT STRAND 86 91 {ECO:0000244|PDB:4P6I}. SQ SEQUENCE 94 AA; 10518 MW; D1C159D924B477B4 CRC64; MSMLVVVTEN VPPRLRGRLA IWLLEVRAGV YVGDVSAKIR EMIWEQIAGL AEEGNVVMAW ATNTETGFEF QTFGLNRRTP VDLDGLRLVS FLPV //