Crystallization and preliminary X‐ray diffraction analysis of the Cmr2–Cmr3 subcomplex in the CRISPR–Cas RNA‐silencing effector complex. Issue 5 (22nd May 2013)

• Record Type: Journal Article
• Authors: Osawa, Takuo; Inanaga, Hideko; Numata, Tomoyuki
• Is Part Of: Acta crystallographica. Volume 69:Issue 5(2013:May)
• Page Start: 585
• Page End: 587
• Publication Date: 2013-05-22
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S1744309113011202
• Languages: English
• ISSNs: 1744-3091

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Clustered, regularly interspaced, short palindromic repeat (CRISPR) loci, found in prokaryotes, are transcribed to produce CRISPR RNAs (crRNAs). The Cmr proteins (Cmr1–6) and crRNA form a ribonucleoprotein complex that degrades target RNAs derived from invading genetic elements. Cmr2dHD, a Cmr2 variant lacking the N‐terminal putative HD nuclease domain, and Cmr3 were co‐expressed in <italic>Escherichia coli</italic> cells and co‐purified as a complex. The Cmr2dHD–Cmr3 complex was co‐crystallized with 3′‐AMP by the vapour‐diffusion method. The crystals diffracted to 2.6 Å resolution using synchrotron radiation at the Photon Factory. The crystals belonged to the orthorhombic space group <italic>I</italic>222, with unit‐cell parameters <italic>a</italic> = 103.9, <italic>b</italic> = 136.7, <italic>c</italic> = 192.0 Å. The asymmetric unit of the crystals is expected to contain one Cmr2dHD–Cmr3 complex with a Matthews coefficient of 3.0 ų Da⁻¹ and a solvent content of 59%.</p> </abstract>