4OUN

Crystal Structure of Mini-ribonuclease 3 from Bacillus subtilis

Summary for 4OUN

• Entry DOI: 10.2210/pdb4oun/pdb
• Descriptor: Mini-ribonuclease 3 (2 entities in total)
• Functional Keywords: rnase iii domain-like, ribonuclease, rna binding, hydrolase
• Biological source: Bacillus subtilis subsp. subtilis
• Total number of polymer chains: 1
• Total formula weight: 18422.74

Authors

Chojnowski, G.,Czarnecka, J.,Nowak, E.,Pianka, D.,Glow, D.,Sabala, I.,Skowronek, K.,Nowotny, M.,Bujnicki, J.M. (deposition date: 2014-02-18, release date: 2015-02-04, Last modification date: 2023-11-08)

Primary citation

Glow, D.,Pianka, D.,Sulej, A.A.,Kozlowski, L.P.,Czarnecka, J.,Chojnowski, G.,Skowronek, K.J.,Bujnicki, J.M.
Sequence-specific cleavage of dsRNA by Mini-III RNase
Nucleic Acids Res., 43:2864-2873, 2015

PubMed Abstract: Ribonucleases (RNases) play a critical role in RNA processing and degradation by hydrolyzing phosphodiester bonds (exo- or endonucleolytically). Many RNases that cut RNA internally exhibit substrate specificity, but their target sites are usually limited to one or a few specific nucleotides in single-stranded RNA and often in a context of a particular three-dimensional structure of the substrate. Thus far, no RNase counterparts of restriction enzymes have been identified which could cleave double-stranded RNA (dsRNA) in a sequence-specific manner. Here, we present evidence for a sequence-dependent cleavage of long dsRNA by RNase Mini-III from Bacillus subtilis (BsMiniIII). Analysis of the sites cleaved by this enzyme in limited digest of bacteriophage Φ6 dsRNA led to the identification of a consensus target sequence. We defined nucleotide residues within the preferred cleavage site that affected the efficiency of the cleavage and were essential for the discrimination of cleavable versus non-cleavable dsRNA sequences. We have also determined that the loop α5b-α6, a distinctive structural element in Mini-III RNases, is crucial for the specific cleavage, but not for dsRNA binding. Our results suggest that BsMiniIII may serve as a prototype of a sequence-specific dsRNase that could possibly be used for targeted cleavage of dsRNA.

PubMed: 25634891
DOI: 10.1093/nar/gkv009

Experimental method

X-RAY DIFFRACTION (1.8 Å)