PDBsum entry 4zt9

Hydrolase/RNA

PDB id: 4zt9

Contents

Protein chains

1335 a.a.

1160 a.a.

DNA/RNA

PDB id:

4zt9

Name:

Hydrolase/RNA

Title:

Nuclease-inactive streptococcus pyogenes cas9 (d10a/h840a, dcas9) in complex with single-guide RNA at 3.1 angstrom resolution

Structure:

Crispr-associated endonuclease cas9. Chain: a, c. Synonym: cas9, crispr-cas9. Engineered: yes. Mutation: yes. Single-guide RNA. Chain: b, d. Engineered: yes

Source:

Streptococcus pyogenes. Organism_taxid: 1314. Gene: csn1, cas9, ers445054_00848, mtb314_0777. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

3.10Å R-factor: 0.245 R-free: 0.284

Authors:

F.Jiang,J.A.Doudna

Key ref:

F.Jiang et al. (2015). STRUCTURAL BIOLOGY. A Cas9-guide RNA complex preorganized for target DNA recognition. Science, 348, 1477-1481. PubMed id: 26113724 DOI: 10.1126/science.aab1452

Date:

14-May-15 Release date: 08-Jul-15

Protein chain

Q99ZW2  (CAS9_STRP1) -  CRISPR-associated endonuclease Cas9/Csn1 from Streptococcus pyogenes serotype M1

Seq: 1368 a.a.

Struc: 1335 a.a.*

Protein chain

Q99ZW2  (CAS9_STRP1) -  CRISPR-associated endonuclease Cas9/Csn1 from Streptococcus pyogenes serotype M1

Seq: 1368 a.a.

Struc: 1160 a.a.*

* PDB and UniProt seqs differ at 6 residue positions (black crosses)

DNA/RNA chains

G-A-U-G-A-G-A-C-G-C-G-U-U-U-U-A-G-A-G-C-U-A-G-A-A-A-U-A-G-C-A-A-G-U-U-A-A-A-A- 72 bases

G-A-U-G-A-G-A-C-G-C-G-U-U-U-U-A-G-A-G-C-U-A-G-A-A-A-U-A-G-C-A-A-G-U-U-A-A-A-A- 72 bases

Enzyme reactions

• Enzyme class: Chains A, C: E.C.3.1.-.-

DOI no: 10.1126/science.aab1452

Science 348:1477-1481 (2015)

PubMed id: 26113724

STRUCTURAL BIOLOGY. A Cas9-guide RNA complex preorganized for target DNA recognition.

F.Jiang, K.Zhou, L.Ma, S.Gressel, J.A.Doudna.

ABSTRACT

Bacterial adaptive immunity uses CRISPR (clustered regularly interspaced short palindromic repeats)-associated (Cas) proteins together with CRISPR transcripts for foreign DNA degradation. In type II CRISPR-Cas systems, activation of Cas9 endonuclease for DNA recognition upon guide RNA binding occurs by an unknown mechanism. Crystal structures of Cas9 bound to single-guide RNA reveal a conformation distinct from both the apo and DNA-bound states, in which the 10-nucleotide RNA "seed" sequence required for initial DNA interrogation is preordered in an A-form conformation. This segment of the guide RNA is essential for Cas9 to form a DNA recognition-competent structure that is poised to engage double-stranded DNA target sequences. We construe this as convergent evolution of a "seed" mechanism reminiscent of that used by Argonaute proteins during RNA interference in eukaryotes.