UniProt UniProt http://www.uniprot.org Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Nature communications 2019 imex curation Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. amunts@scilifelab.se,saikrishnan@iiserpune.ac.in 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro 3d-em 3D electron microscopy 3D-EM electron tomog structure determination identification by structure determination identification by structure determination structure determination Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. amunts@scilifelab.se,saikrishnan@iiserpune.ac.in Accepted 2020-AUG-24 AT 11:38 BST AT 11:38 BST by PPORRAS Nature communications 2019 Only protein-protein interactions imex curation imex curation Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Nature communications 2019 imex curation Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. amunts@scilifelab.se,saikrishnan@iiserpune.ac.in 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. amunts@scilifelab.se,saikrishnan@iiserpune.ac.in Accepted 2020-AUG-24 AT 11:38 BST AT 11:38 BST by PPORRAS Nature communications 2019 Only protein-protein interactions imex curation imex curation Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Nature communications 2019 imex curation Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. amunts@scilifelab.se,saikrishnan@iiserpune.ac.in 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro nuclease assay nuclease assay predetermined predetermined participant predetermined Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. amunts@scilifelab.se,saikrishnan@iiserpune.ac.in Accepted 2020-AUG-24 AT 11:38 BST AT 11:38 BST by PPORRAS Nature communications 2019 Only protein-protein interactions imex curation imex curation Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Nature communications 2019 imex curation Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. amunts@scilifelab.se,saikrishnan@iiserpune.ac.in 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro fluorescence fluorescence technology fluorescence tag fluorescence tag visualisation by fluorescence tag fluorescence Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. Nucleotide binding by mcrB was assessed using the time-dependent change in fluorescence of 2’/3’-O-(N-methyl-anthraniloyl) GNP (mantGNP) amunts@scilifelab.se,saikrishnan@iiserpune.ac.in Accepted 2020-AUG-24 AT 11:38 BST AT 11:38 BST by PPORRAS Nature communications 2019 Only protein-protein interactions imex curation imex curation Mock publication and experiment for abstract interactions that are not interaction evidences. Mock publication and experiment for abstract interactions that are not interaction evidences. unspecified method Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Structure-based mechanism for activation of the AAA+ GTPase McrB by the endonuclease McrC. Nature communications 2019 imex curation Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. amunts@scilifelab.se,saikrishnan@iiserpune.ac.in 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro gtpase assay gtpase assay GTPase gtp hydrolisis gtp hydrolisis GTPase predetermined predetermined participant predetermined Nirwan N., Itoh Y., Singh P., Bandyopadhyay S., Vinothkumar KR., Amunts A., Saikrishnan K. amunts@scilifelab.se,saikrishnan@iiserpune.ac.in Accepted 2020-AUG-24 AT 11:38 BST AT 11:38 BST by PPORRAS Nature communications 2019 Only protein-protein interactions imex curation imex curation gtp guanosine triphosphate - GTP 5'-GTP GTP Guanosine 5'-triphosphate guanosine 5'-triphosphoric acid guanosine triphosphate GUANOSINE-5'-TRIPHOSPHATE H4gtp guanosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 guanosine 5'-[beta,gamma-imido]triphosphate guanosine 5'-[beta,gamma-imido]triphosphate 5'-Guanylic acid, monoanhydride with imidodiphosphoric acid 5'-guanylylimidodiphosphate GMP-PNP GMP-Pnp Gpp(NH)-p Gpp(NH)p Guanylyl imidodiphosphate 5'-O-(hydroxy{[hydroxy(phosphonoamino)phosphoryl]oxy}phosphoryl)guanosine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) mcrc_ecoli Protein McrC mcrC b4345 JW5789 protein protein ecoli Escherichia coli (strain K12) MEQPVIPVRNIYYMLTYAWGYLQEIKQANLEAIPGNNLLDILGYVLNKGVLQLSRRGLELDYNPNTEIIPGIKGRIEFAKTIRGFHLNHGKTVSTFDMLNEDTLANRIIKSTLAILIKHEKLNSTIRDEARSLYRKLPGISTLHLTPQHFSYLNGGKNTRYYKFVISVCKFIVNNSIPGQNKGHYRFYDFERNEKEMSLLYQKFLYEFCRRELTSANTTRSYLKWDASSISDQSLNLLPRMETDITIRSSEKILIVDAKYYKSIFSRRMGTEKFHSQNLYQLMNYLWSLKPENGENIGGLLIYPHVDTAVKHRYKINGFDIGLCTVNLGQEWPCIHQELLDIFDEYLK 9B65FAD20E5C6E64 mcrb_dna-5prime McrC nuclease substrate dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid chemical synthesis Chemical synthesis (Chemical synthesis) TAATACGACTCACTATAGGG mcrb_dna-3prime McrB nuclease substrate dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid chemical synthesis Chemical synthesis (Chemical synthesis) GCTAGTTATTGCTCAGCGG mcrb_ecoli 5-methylcytosine-specific restriction enzyme B mcrB rglB b4346 EcoKMcrBC JW5871 protein protein ecoli Escherichia coli (strain K12) MESIQPWIEKFIKQAQQQRSQSTKDYPTSYRNLRVKLSFGYGNFTSIPWFAFLGEGQEASNGIYPVILYYKDFDELVLAYGISDTNEPHAQWQFSSDIPKTIAEYFQATSGVYPKKYGQSYYACSQKVSQGIDYTRFASMLDNIINDYKLIFNSGKSVIPPMSKTESYCLEDALNDLFIPETTIETILKRLTIKKNIILQGPPGVGKTFVARRLAYLLTGEKAPQRVNMVQFHQSYSYEDFIQGYRPNGVGFRRKDGIFYNFCQQAKEQPEKKYIFIIDEINRANLSKVFGEVMMLMEHDKRGENWSVPLTYSENDEERFYVPENVYIIGLMNTADRSLAVVDYALRRRFSFIDIEPGFDTPQFRNFLLNKKAEPSFVESLCQKMNELNQEISKEATILGKGFRIGHSYFCCGLEDGTSPDTQWLNEIVMTDIAPLLEEYFFDDPYKQQKWTNKLLGDS DIP protein P15005 original sequence version: 91 DIP protein P15005 original sequence version: null C8167978B0D720CA mcrb-mcrc-1 1 9 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role neutral component neutral component purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Supp. Table 2 mcrb-mcrb-1 2 12 unspecified role unspecified role neutral component neutral component purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P15005:p.Arg348Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 20:14:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg348Ala' P15005:p.Arg349Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 16:53:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg349Ala' P15005:p.Glu298Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain E A Tue Jun 30 19:40:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu298Ala' ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role neutral component neutral component purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P15005:p.Arg348Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 18:23:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg348Ala' P15005:p.Arg349Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 19:46:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg349Ala' P15005:p.Glu298Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain E A Tue Jun 30 17:47:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu298Ala' ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 2G mcrb-mcrc-2 2 9 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role neutral component neutral component purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P15005:p.Arg348Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 18:33:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg348Ala' P15005:p.Arg349Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 17:54:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg349Ala' P15005:p.Glu298Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain E A Tue Jun 30 19:35:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu298Ala' ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 2G (inset) Interaction occurs in the presence of GTP guanosine 5'_[beta,gamma_imido]triphosphate-mcrb-1 4 8 unspecified role unspecified role neutral component neutral component region fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role neutral component neutral component purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P15005:p.Arg348Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 17:45:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg348Ala' P15005:p.Arg349Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 18:04:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg349Ala' P15005:p.Glu298Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain E A Tue Jun 30 18:34:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu298Ala' ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 2H gtp-mcrb_mcrc_ecoli-1 6 50 enzyme enzyme neutral component neutral component purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P15005:p.Arg348Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain R A 54 Mutation update was not able to retrieve the sequence, please check Kindly select the protein which had been mutated P15005:p.Arg349Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain R A 54 Mutation update was not able to retrieve the sequence, please check kindly select the mutated participant P15005:p.Glu298Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain E A 54 Mutation update was not able to retrieve the sequence, please check Kindly select the protein which had been mutated ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component region fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined gtpase reaction gtpase reaction Fig. 2I Kindly assign the mutations to respective participants gtp-mcrb_mcrc_ecoli-2 3 50 enzyme enzyme neutral component neutral component purified purified c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P15005:p.Arg348Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain R A 54 Mutation update was not able to retrieve the sequence, please check Kindly select the protein which had been mutated P15005:p.Arg349Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain R A 54 Mutation update was not able to retrieve the sequence, please check kindly select the mutated participant in the complex.. P15005:p.Glu298Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain E A 54 Mutation update was not able to retrieve the sequence, please check Kindly select the protein which had been mutated ecoli Escherichia coli (strain K12) 11 enzyme target enzyme target substrate substrate unspecified role unspecified role region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 enzyme target enzyme target substrate substrate neutral component neutral component region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 69 67 gtpase reaction gtpase reaction Fig. 2J Interaction occurs in the presence of GTP Kindly assign mutations to respective participants mcrbc_ecoli McrBC 5-methylcytosine-specific restriction endonuclease complex 12xmcrB:2xmcrC 9 unspecified role unspecified role mcrb binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined mcrc binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 enzyme enzyme mcrc binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role mcrb binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined mcrc binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 enzyme enzyme mcrc binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 71 74 72 77 76 78 physical association physical association ecoli Escherichia coli (strain K12) stable complex stable complex physical interaction evidence used in manual assertion physical interaction evidence used in manual assertion IPI inferred from physical interaction A DNA endonuclease that acts as an antiphage defense system by specifically cleaving invading DNA that are methylated. McrBC specifically cleaves DNA containing the recognition sequence 5prime-G/A(5mC)-3prime, where 5mC can be 5-methylcytosine, 5-hydroxymethylcytosine, or 4-methylcytosine. The AAA+ GTPase motor mcrB is activated by mcrC, which in turn powers DNA cleavage by the endonuclease. Two hexameric rings of mcrB are bridged by an mcrC dimer. The inside of the central channel of the AAA+ mcrB ring shows a circular patch of positively charged residues, suggesting that DNA threads through the central channel. Heterotetradecamer Accepted 2020-MAR-31 by LIVIAP