Abstract
Binase is an extracellular guanyl-preferring ribonuclease from Bacillus pumilus. The main biological function of binase is RNA degradation with the formation of guanosine-2',3'-cyclic phosphate and its subsequent hydrolysis to 3'-phosphate. Extracellular RNases are believed to be key agents that affect the functional activity of the body, as they directly interact with epithelial and immune cells. The biological effects of the enzyme may consist of both direct RNA degradation, and the accumulation of 2',3'-cGMP in the human body. In this work, we have performed a comparative analysis of the cleavage efficiency of model RNA substrates, i.e., short hairpin structures that contain guanosine at various positions. It has been shown that the hydrolysis efficiency of the model RNA substrates depends on the position of guanosine. We have also demonstrated the influence of various divalent metal ions and low molecular weight nucleotide compounds on the binase-catalyzed endoribonucleolytic reaction.
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Funding
The work was supported by grant no. 19-74-10034 from the Russian Science Foundation (the design and synthesis of RNA substrates, analysis of the enzymatic process by gel electrophoresis and stop flow method), grant no. 17-00-00060 from the Russian Foundation for Basic Research, partially project no. АААА-А17-117020210022-4 from the Russian State-funded budget (the use of the equipment), and the program for improving competitiveness of Kazan (Volga Region) Federal University.
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Translated by A. Levina
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Kuznetsova, A.A., Akhmetgalieva, A.A., Ulyanova, V.V. et al. Efficiency of RNA Hydrolysis by Binase from Bacillus pumilus: The Impact of Substrate Structure, Metal Ions, and Low Molecular Weight Nucleotide Compounds. Mol Biol 54, 769–776 (2020). https://doi.org/10.1134/S0026893320050064
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DOI: https://doi.org/10.1134/S0026893320050064