title: Purification and crystallization of Bacillus subtilis NrnA, a novel enzyme involved in nanoRNA degradation.
abstract: The final step in RNA degradation is the hydrolysis of RNA fragments five nucleotides or less in length (nanoRNA) to mononucleotides. In Escherichia coli this step is carried out by oligoribonuclease (Orn), a DEDD-family exoribonuclease that is conserved throughout eukaryotes. However, many bacteria lack Orn homologs, and an unrelated DHH-family phosphoesterase, NrnA, has recently been identified as one of the enzymes responsible for nanoRNA degradation in Bacillus subtilis. To understand its mechanism of action, B. subtilis NrnA was purified and crystallized at room temperature using the hanging-drop vapor-diffusion method with PEG 4000, PEG 3350 or PEG MME 2000 as precipitant. The crystals belonged to the primitive monoclinic space group P2(1), with unit-cell parameters a = 50.62, b = 121.3, c = 123.4 Å, α = 90, β = 91.31, γ = 90°.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, PO Box 016129, Miami, FL 33101-6129, USA.
authors.forename: Claudiu M
authors.initials: CM
authors.lastname: Nelersa
authors.forename: Brad J
authors.initials: BJ
authors.lastname: Schmier
authors.forename: Arun
authors.initials: A
authors.lastname: Malhotra
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D009708:Nucleotidases / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D009708:Nucleotidases; C053354:bisphosphoadenylate 3'-nucleotidase
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