EC Number   |
Substrates |
Organism |
Products |
Reversibility |
|---|
   3.1.12.1 | more |
the enzyme also exhibits ATP-independent DNA unwinding activity |
Saccharolobus solfataricus |
? |
- |
? |
| 3.1.12.1 | more |
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 |
? |
- |
? |
| 3.1.12.1 | more |
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 | more |
Cas4 proteins contain a RecB-like nuclease domain and exhibit in vitro exonuclease activities |
Synechocystis sp. PCC 6803 |
? |
- |
? |
| 3.1.12.1 | more |
Cas4 proteins contain a RecB-like nuclease domain and exhibit in vitro exonuclease activities |
Pyrococcus furiosus |
? |
- |
? |
| 3.1.12.1 | more |
Cas4 proteins contain a RecB-like nuclease domain and exhibit in vitro exonuclease activities |
Escherichia coli |
? |
- |
? |
| 3.1.12.1 | more |
Cas4 proteins contain a RecB-like nuclease domain and exhibit in vitro exonuclease activities. In the presence of Cas1 and Cas2, Cas4 of Bacillus halodurans endonucleolytically cuts long 3' overhangs of prespacers at the PAM site |
Halalkalibacterium halodurans |
? |
- |
? |
| 3.1.12.1 | more |
Cas4-1 trims and orients a duplexed DNA oligonucleotide pre-spacer. Nuclease |
Pyrococcus furiosus |
? |
- |
? |
| 3.1.12.1 | more |
Cas4-2 trims and orients a duplexed DNA oligonucleotide pre-spacer |
Pyrococcus furiosus |
? |
- |
? |
| 3.1.12.1 | more |
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 |
? |
- |
? |
