5wtk

Structural highlights

Function

CS13A_LEPSD CRISPR (clustered regularly interspaced short palindromic repeat), is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain sequences complementary to antecedent mobile elements (spacers) and target invading nucleic acids. Unlike many single-component effectors, this CRISPR-Cas system targets RNA (PubMed:27256883, PubMed:28086085). CRISPR clusters are transcribed from pre-CRISPR RNA (crRNA) and processed into crRNA (optimally 28 nucleotides in this system) by this protein (PubMed:27256883, PubMed:27669025, PubMed:28475872, PubMed:28086085). This protein processes pre-crRNA at a 'non-typical' site 1 nucleotide upstream of the pre-crRNA stem-loop; it cleaves pre-crRNA from L.buccalis and L.wadei in a similar fashion, whereas the enzymes from the latter 2 bacteria cleave their own pre-crRNA 3 nt further upstream (PubMed:28475872). When the appropriate target sequences are cloned into the CRISPR array, confers immunity to ssRNA(+) enterobacteria phage MS2 (PubMed:27256883). Cleaves linear target ssRNA in a crRNA-dependent fashion, preferentially before U residues; has no activity on partially dsRNA, ssDNA or dsDNA (PubMed:27256883). RNA secondary structure surrounding the target influence the cleavage site and efficiency; unlike other CRISPR-Cas effectors Cas13a cleaves outside of the crRNA binding site (PubMed:27256883). In the presence of a viable RNA target other RNAs are also degraded (called collateral RNA degradation), suggesting this type of CRISPR-Cas might also prevent viral spread by inducing programmed cell death or dormancy (PubMed:27256883). This system has a 3' protospacer flanking site (PFS), it does not cleave when the 3' PFS is G (PFS is equivalent to PAM, the protospacer adjacent motif) (PubMed:27256883). Mutations of its active site residues results in an RNA-programmed RNA-binding protein (PubMed:27256883).^{[1] [2] [3] [4]}

References

1. ↑ Abudayyeh OO, Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox DB, Shmakov S, Makarova KS, Semenova E, Minakhin L, Severinov K, Regev A, Lander ES, Koonin EV, Zhang F. C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Aug 5;353(6299):aaf5573. doi: 10.1126/science.aaf5573. Epub 2016, Jun 2. PMID:27256883 doi:http://dx.doi.org/10.1126/science.aaf5573
2. ↑ East-Seletsky A, O'Connell MR, Knight SC, Burstein D, Cate JH, Tjian R, Doudna JA. Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Oct 13;538(7624):270-273. doi: 10.1038/nature19802. Epub 2016 Sep, 26. PMID:27669025 doi:http://dx.doi.org/10.1038/nature19802
3. ↑ Liu L, Li X, Wang J, Wang M, Chen P, Yin M, Li J, Sheng G, Wang Y. Two Distant Catalytic Sites Are Responsible for C2c2 RNase Activities. Cell. 2017 Jan 12;168(1-2):121-134.e12. doi: 10.1016/j.cell.2016.12.031. Epub, 2017 Jan 12. PMID:28086085 doi:http://dx.doi.org/10.1016/j.cell.2016.12.031
4. ↑ East-Seletsky A, O'Connell MR, Burstein D, Knott GJ, Doudna JA. RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. PMID:28475872 doi:10.1016/j.molcel.2017.04.008
5. ↑ Liu L, Li X, Wang J, Wang M, Chen P, Yin M, Li J, Sheng G, Wang Y. Two Distant Catalytic Sites Are Responsible for C2c2 RNase Activities. Cell. 2017 Jan 12;168(1-2):121-134.e12. doi: 10.1016/j.cell.2016.12.031. Epub, 2017 Jan 12. PMID:28086085 doi:http://dx.doi.org/10.1016/j.cell.2016.12.031