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Literature summary for 3.1.12.1 extracted from
- Cas4-dependent prespacer processing ensures high-fidelity programming of CRISPR arrays (2018), Mol. Cell., 70, 48-59.e5 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene BH0340, recombinant expression of His-tagged enzyme Cas4 in Escherichia coli | Halalkalibacterium halodurans |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Halalkalibacterium halodurans | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | ? | - |
? |  |
| additional information | Halalkalibacterium halodurans ATCC BAA-125 | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | ? | - |
? | |
| additional information | Halalkalibacterium halodurans JCM 9153 | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | ? | - |
? | |
| additional information | Halalkalibacterium halodurans FERM 7344 | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | ? | - |
? | |
| additional information | Halalkalibacterium halodurans C-125 | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | ? | - |
? | |
| additional information | Halalkalibacterium halodurans DSM 18197 | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Halalkalibacterium halodurans | Q9KFY0 | - |
- |
| Halalkalibacterium halodurans ATCC BAA-125 | Q9KFY0 | - |
- |
| Halalkalibacterium halodurans C-125 | Q9KFY0 | - |
- |
| Halalkalibacterium halodurans DSM 18197 | Q9KFY0 | - |
- |
| Halalkalibacterium halodurans FERM 7344 | Q9KFY0 | - |
- |
| Halalkalibacterium halodurans JCM 9153 | Q9KFY0 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme Cas4 from Escherichia coli by nickel affinity chromatography, co-elution of a Cas4-Cas1-Cas2 complex from nickel affinity purification | Halalkalibacterium halodurans |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | Halalkalibacterium halodurans | ? | - |
? | |
| additional information | in the presence of Cas1 and Cas2, Cas4 processes double-stranded substrates with long 3' overhangs through site-specific endonucleolytic cleavage. Cas4 recognizes PAM sequences within the prespacer and prevents integration of unprocessed prespacers, ensuring that only functional spacers will be integrated into the CRISPR array. Assays with dsDNA substrates containing blunt ends or a 24-bp duplex flanked by 15-nt 3' or 5' overhangs result in cleavage of the 3'-overhanging substrates at 65°C, but no cleavage of 5'-overhanging or blunt-end substrates by the enzyme under similar conditions. Exonucleolytic cleavage of the blunt-end substrate is observed using free Cas4, but only at very high concentrations. Sequence-specific integration and asymmetric prespacer processing by the adaptation complex | Halalkalibacterium halodurans | ? | - |
? | |
| additional information | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | Halalkalibacterium halodurans ATCC BAA-125 | ? | - |
? | |
| additional information | in the presence of Cas1 and Cas2, Cas4 processes double-stranded substrates with long 3' overhangs through site-specific endonucleolytic cleavage. Cas4 recognizes PAM sequences within the prespacer and prevents integration of unprocessed prespacers, ensuring that only functional spacers will be integrated into the CRISPR array. Assays with dsDNA substrates containing blunt ends or a 24-bp duplex flanked by 15-nt 3' or 5' overhangs result in cleavage of the 3'-overhanging substrates at 65°C, but no cleavage of 5'-overhanging or blunt-end substrates by the enzyme under similar conditions. Exonucleolytic cleavage of the blunt-end substrate is observed using free Cas4, but only at very high concentrations. Sequence-specific integration and asymmetric prespacer processing by the adaptation complex | Halalkalibacterium halodurans ATCC BAA-125 | ? | - |
? | |
| additional information | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | Halalkalibacterium halodurans JCM 9153 | ? | - |
? | |
| additional information | in the presence of Cas1 and Cas2, Cas4 processes double-stranded substrates with long 3' overhangs through site-specific endonucleolytic cleavage. Cas4 recognizes PAM sequences within the prespacer and prevents integration of unprocessed prespacers, ensuring that only functional spacers will be integrated into the CRISPR array. Assays with dsDNA substrates containing blunt ends or a 24-bp duplex flanked by 15-nt 3' or 5' overhangs result in cleavage of the 3'-overhanging substrates at 65°C, but no cleavage of 5'-overhanging or blunt-end substrates by the enzyme under similar conditions. Exonucleolytic cleavage of the blunt-end substrate is observed using free Cas4, but only at very high concentrations. Sequence-specific integration and asymmetric prespacer processing by the adaptation complex | Halalkalibacterium halodurans JCM 9153 | ? | - |
? | |
| additional information | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | Halalkalibacterium halodurans FERM 7344 | ? | - |
? | |
| additional information | in the presence of Cas1 and Cas2, Cas4 processes double-stranded substrates with long 3' overhangs through site-specific endonucleolytic cleavage. Cas4 recognizes PAM sequences within the prespacer and prevents integration of unprocessed prespacers, ensuring that only functional spacers will be integrated into the CRISPR array. Assays with dsDNA substrates containing blunt ends or a 24-bp duplex flanked by 15-nt 3' or 5' overhangs result in cleavage of the 3'-overhanging substrates at 65°C, but no cleavage of 5'-overhanging or blunt-end substrates by the enzyme under similar conditions. Exonucleolytic cleavage of the blunt-end substrate is observed using free Cas4, but only at very high concentrations. Sequence-specific integration and asymmetric prespacer processing by the adaptation complex | Halalkalibacterium halodurans FERM 7344 | ? | - |
? | |
| additional information | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | Halalkalibacterium halodurans C-125 | ? | - |
? | |
| additional information | in the presence of Cas1 and Cas2, Cas4 processes double-stranded substrates with long 3' overhangs through site-specific endonucleolytic cleavage. Cas4 recognizes PAM sequences within the prespacer and prevents integration of unprocessed prespacers, ensuring that only functional spacers will be integrated into the CRISPR array. Assays with dsDNA substrates containing blunt ends or a 24-bp duplex flanked by 15-nt 3' or 5' overhangs result in cleavage of the 3'-overhanging substrates at 65°C, but no cleavage of 5'-overhanging or blunt-end substrates by the enzyme under similar conditions. Exonucleolytic cleavage of the blunt-end substrate is observed using free Cas4, but only at very high concentrations. Sequence-specific integration and asymmetric prespacer processing by the adaptation complex | Halalkalibacterium halodurans C-125 | ? | - |
? | |
| additional information | Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 enhances prespacer processing. Mechanism and structural and mechanistic model, overview | Halalkalibacterium halodurans DSM 18197 | ? | - |
? | |
| additional information | in the presence of Cas1 and Cas2, Cas4 processes double-stranded substrates with long 3' overhangs through site-specific endonucleolytic cleavage. Cas4 recognizes PAM sequences within the prespacer and prevents integration of unprocessed prespacers, ensuring that only functional spacers will be integrated into the CRISPR array. Assays with dsDNA substrates containing blunt ends or a 24-bp duplex flanked by 15-nt 3' or 5' overhangs result in cleavage of the 3'-overhanging substrates at 65°C, but no cleavage of 5'-overhanging or blunt-end substrates by the enzyme under similar conditions. Exonucleolytic cleavage of the blunt-end substrate is observed using free Cas4, but only at very high concentrations. Sequence-specific integration and asymmetric prespacer processing by the adaptation complex | Halalkalibacterium halodurans DSM 18197 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | Cas4 forms a tight heterohexameric complex with the Cas1 spacer integrase containing two Cas1 dimers and two Cas4 subunits | Halalkalibacterium halodurans |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| BH0340 | - |
Halalkalibacterium halodurans |
| Cas4 | - |
Halalkalibacterium halodurans |
| type I-C Cas4 | - |
Halalkalibacterium halodurans |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 65 | - |
assay at | Halalkalibacterium halodurans |
General Information
| General Information | Comment | Organism |
|---|---|---|
| physiological function | Bacillus halodurans type I-C Cas4 is required for efficient prespacer processing prior to Cas1-Cas2-mediated integration. Cas4, a core family of CRISPR-associated proteins, associates with the Cas1 spacer integrase and is required for efficient prespacer processing during CRISPR-Cas adaptation. Cas4 processing prevents non-functional spacers from being integrated into the CRISPR array, ensuring the fidelity of the adaptation process. Cas4 forms a tight heterohexameric complex with the Cas1 spacer integrase. Mechanism, overview. Cas4 is a Cas1-Cas2-dependent endonuclease that cleaves 3' overhangs of prespacers. Cas4 cleavage is sequence and site-specific and depends on the presence of a protospacer adjacent motif (PAM). Cas4 blocks premature integration of uncleaved prespacers, ensuring spacer fidelity | Halalkalibacterium halodurans |
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