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Literature summary extracted from
- Cas4 nucleases define the PAM, length, and orientation of DNA fragments integrated at CRISPR loci (2018), Mol. Cell., 70, 814-824.e6 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.1.12.1 | gene PF1119 or cas4-1, Cas gene loci and the adaptation assay, the cas4-1 gene is located immediately adjacent to cas1 and cas2 | Pyrococcus furiosus |
| 3.1.12.1 | gene PF1793 or cas4-2, Cas gene loci and the adaptation assay, the cas4-2 gene resides in a remote genomic location | Pyrococcus furiosus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.1.12.1 | additional information | cas4-2 dene deletion | Pyrococcus furiosus |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.12.1 | additional information | Pyrococcus furiosus | Cas4-1 trims and orients a duplexed DNA oligonucleotide pre-spacer. Nuclease activities of Cas4 proteins are essential to define PAM, length, and orientation of new spacers | ? | - |
? |  |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.1.12.1 | Pyrococcus furiosus | Q8U027 | - |
- |
| 3.1.12.1 | Pyrococcus furiosus | Q8U1T6 | - |
- |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.12.1 | additional information | Cas4-1 trims and orients a duplexed DNA oligonucleotide pre-spacer. Nuclease activities of Cas4 proteins are essential to define PAM, length, and orientation of new spacers | Pyrococcus furiosus | ? | - |
? | |
| 3.1.12.1 | additional information | Cas4-1 trims and orients a duplexed DNA oligonucleotide pre-spacer. Nuclease | Pyrococcus furiosus | ? | - |
? | |
| 3.1.12.1 | additional information | Cas4-2 trims and orients a duplexed DNA oligonucleotide pre-spacer | Pyrococcus furiosus | ? | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.1.12.1 | Cas4 | - |
Pyrococcus furiosus |
| 3.1.12.1 | Cas4 nuclease | - |
Pyrococcus furiosus |
| 3.1.12.1 | Cas4-1 | - |
Pyrococcus furiosus |
| 3.1.12.1 | Cas4-2 | - |
Pyrococcus furiosus |
| 3.1.12.1 | PF1119 | - |
Pyrococcus furiosus |
| 3.1.12.1 | PF1793 | - |
Pyrococcus furiosus |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 3.1.12.1 | malfunction | while cas4-2 deletion does not increase adaptation frequency overall, it dramatically increases incorporation of the duplexed DNA oligonucleotide. The plasmid-expressed wild-type Cas4-2 rescues spacer sizes and the downstream NW motif, but the nuclease-defective Cas4-2 does not | Pyrococcus furiosus |
| 3.1.12.1 | additional information | analysis of protospace adjacent motif (PAM) sequence recognition by the Cas4 isozymes, overview | Pyrococcus furiosus |
| 3.1.12.1 | physiological function | two distinct Cas4 nucleases are essential for CRISPR DNA acquisition in Pyrococcus furiosus. Cas4-1 defines the 5' NGG PAM, and Cas4-2 defines the 3' NW motif. Cas4-1 and Cas4-2 nucleases trim opposite ends of prespacer DNA to the correct size. Both Cas4 proteins direct integration of DNA in the correct orientation for immunity. The concerted action of two different Cas4 nucleases is required for protospace adjacent motif (PAM) definition, prespacer processing, and integration of DNA fragments at the CRISPR array in a functional orientation. Pyrococcus furiosus actively takes up spacers into each CRISPR locus in a PAM-dependent manner. Adaptation occurs under normal growth conditions, and the frequency of adaptation can be increased by overexpression of Cas1, Cas2, and Cas4 proteins. Cas4-1 and Cas4-2 nuclease activity is essential for PAM recognition and spacer sizing | Pyrococcus furiosus |
| 3.1.12.1 | physiological function | two distinct Cas4 nucleases are essential for CRISPR DNA acquisition in Pyrococcus furiosus. Cas4-1 defines the 5' NGG PAM, and Cas4-2 defines the 3' NW motif. Cas4-1 and Cas4-2 nucleases trim opposite ends of prespacer DNA to the correct size. Both Cas4 proteins direct integration of DNA in the correct orientation for immunity. The concerted action of two different Cas4 nucleases is required for protospacer adjacent motif (PAM) definition, prespacer processing, and integration of DNA fragments at the CRISPR array in a functional orientation. Pyrococcus furiosus actively takes up spacers into each CRISPR locus in a PAM-dependent manner. Adaptation occurs under normal growth conditions, and the frequency of adaptation can be increased by overexpression of Cas1, Cas2, and Cas4 proteins. Cas4-1 and Cas4-2 nuclease activity is essential for PAM recognition and spacer sizing | Pyrococcus furiosus |
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