PDBsum entry 4oge

Hydrolase

PDB id: 4oge

Contents

Protein chain

977 a.a.

Ligands

SPD ×2

Metals

_ZN

_MG ×3

Waters ×343

PDB id:

4oge

Name:

Hydrolase

Title:

Crystal structure of the type ii-c cas9 enzyme from actinomyces naeslundii

Structure:

Hnh endonuclease domain protein. Chain: a. Engineered: yes

Source:

Actinomyces naeslundii. Organism_taxid: 1115803. Strain: howell 279. Gene: hmpref1129_2620. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.20Å R-factor: 0.188 R-free: 0.226

Authors:

F.Jiang,E.Ma,S.Lin,J.A.Doudna

Key ref:

M.Jinek et al. (2014). Structures of Cas9 endonucleases reveal RNA-mediated conformational activation. Science, 343, 1247997. PubMed id: 24505130 DOI: 10.1126/science.1247997

Date:

15-Jan-14 Release date: 12-Feb-14

Protein chain

J3F2B0  (CAS9_ACTNH) -  CRISPR-associated endonuclease Cas9 from Actinomyces naeslundii (strain ATCC 12104 / DSM 43013 / CCUG 2238 / JCM 8349 / NCTC 10301 / Howell 279)

Seq: 1101 a.a.

Struc: 977 a.a.

Enzyme reactions

• Enzyme class: E.C.3.1.-.-

DOI no: 10.1126/science.1247997

Science 343:1247997 (2014)

PubMed id: 24505130

Structures of Cas9 endonucleases reveal RNA-mediated conformational activation.

M.Jinek, F.Jiang, D.W.Taylor, S.H.Sternberg, E.Kaya, E.Ma, C.Anders, M.Hauer, K.Zhou, S.Lin, M.Kaplan, A.T.Iavarone, E.Charpentier, E.Nogales, J.A.Doudna.

ABSTRACT

Type II CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems use an RNA-guided DNA endonuclease, Cas9, to generate double-strand breaks in invasive DNA during an adaptive bacterial immune response. Cas9 has been harnessed as a powerful tool for genome editing and gene regulation in many eukaryotic organisms. We report 2.6 and 2.2 angstrom resolution crystal structures of two major Cas9 enzyme subtypes, revealing the structural core shared by all Cas9 family members. The architectures of Cas9 enzymes define nucleic acid binding clefts, and single-particle electron microscopy reconstructions show that the two structural lobes harboring these clefts undergo guide RNA-induced reorientation to form a central channel where DNA substrates are bound. The observation that extensive structural rearrangements occur before target DNA duplex binding implicates guide RNA loading as a key step in Cas9 activation.