3S4L

The CRISPR-associated Cas3 HD domain protein MJ0384 from Methanocaldococcus jannaschii

Replaces:  3M5F

Summary for 3S4L

• Entry DOI: 10.2210/pdb3s4l/pdb
• Descriptor: CAS3 Metal dependent phosphohydrolase, CALCIUM ION (3 entities in total)
• Functional Keywords: immune system, hd-motif, structural genomics, psi-biology, midwest center for structural genomics, mcsg, nuclease, hydrolase
• Biological source: Methanocaldococcus jannaschii
• Total number of polymer chains: 1
• Total formula weight: 28532.87

Authors

Petit, P.,Brown, G.,Yakunin, A.,Edwards, A.,Joachimiak, A.,Savchenko, A.,Midwest Center for Structural Genomics (MCSG) (deposition date: 2011-05-19, release date: 2011-06-22, Last modification date: 2024-02-28)

Primary citation

Beloglazova, N.,Petit, P.,Flick, R.,Brown, G.,Savchenko, A.,Yakunin, A.F.
Structure and activity of the Cas3 HD nuclease MJ0384, an effector enzyme of the CRISPR interference.
Embo J., 30:4616-4627, 2011

PubMed Abstract: Clustered regularly interspaced short palindromic repeats (CRISPRs) and Cas proteins represent an adaptive microbial immunity system against viruses and plasmids. Cas3 proteins have been proposed to play a key role in the CRISPR mechanism through the direct cleavage of invasive DNA. Here, we show that the Cas3 HD domain protein MJ0384 from Methanocaldococcus jannaschii cleaves endonucleolytically and exonucleolytically (3'-5') single-stranded DNAs and RNAs, as well as 3'-flaps, splayed arms, and R-loops. The degradation of branched DNA substrates by MJ0384 is stimulated by the Cas3 helicase MJ0383 and ATP. The crystal structure of MJ0384 revealed the active site with two bound metal cations and together with site-directed mutagenesis suggested a catalytic mechanism. Our studies suggest that the Cas3 HD nucleases working together with the Cas3 helicases can completely degrade invasive DNAs through the combination of endo- and exonuclease activities.

PubMed: 22009198
DOI: 10.1038/emboj.2011.377

Experimental method

X-RAY DIFFRACTION (2.3 Å)