Open AccessPurification and crystallization of Bacillus subtilis NrnA, a novel enzyme involved in nanoRNA degradation
Author(s) -
Nelersa Claudiu M.,
Schmier Brad J.,
Malhotra Arun
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111026509
Subject(s) - bacillus subtilis , escherichia coli , enzyme , exoribonuclease , peg ratio , degradation (telecommunications) , rna , crystallization , biology , chemistry , recombinant dna , bacteria , biochemistry , genetics , gene , organic chemistry , telecommunications , finance , economics , computer science , rnase p
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 P 2 1 , with unit‐cell parameters a = 50.62, b = 121.3, c = 123.4 Å, α = 90, β = 91.31, γ = 90°.