Purification and crystallization of Bacillus subtilis NrnA, a novel enzyme involved in nanoRNA degradation

Journal Article · Tue May 08 04:00:00 EDT 2012 · Acta Crystallogr. F

DOI:https://doi.org/10.1107/S1744309111026509· OSTI ID:1036028

Nelersa, Claudiu M; Schmier, Brad J; Malhotra, Arun ^[1]

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1. Miami-MED

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{sub 1}, with unit-cell parameters a = 50.62, b = 121.3, c = 123.4 {angstrom}, {alpha} = 90, {beta} = 91.31, {gamma} = 90{sup o}.

• Research Organization: Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)
• Sponsoring Organization: AHANIH
• OSTI ID: 1036028
• Journal Information: Acta Crystallogr. F, Journal Name: Acta Crystallogr. F Journal Issue: (10) ; 10, 2011 Vol. 67; ISSN 1744-3091
• Country of Publication: United States
• Language: ENGLISH

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
BACILLUS SUBTILIS
BACTERIA
CRYSTALLIZATION
ENZYMES
ESCHERICHIA COLI
HYDROLYSIS
NUCLEOTIDES
PURIFICATION
RNA
SPACE GROUPS