NDLI: Effects of background anionic compounds on the activity of the hammerhead ribozyme in $Mg^{2+}-unsaturated$ solutions

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Effects of background anionic compounds on the activity of the hammerhead ribozyme in $Mg^{2+}-unsaturated$ solutions

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Nakano, Shu-ichi Kitagawa, Yuichi Miyoshi, Daisuke Sugimoto, Naoki
Description	Author Affiliation: Nakano S ( Department of Nanobiochemistry, Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, 7-1-20, Minatojima-minamimachi, Chuo-ku, Kobe, 650-0047, Japan, shuichi@center.konan-u.ac.jp.)
Abstract	Cellular ribozymes exhibit catalytic activity in media containing large numbers of anionic compounds and macromolecules. In this study, the RNA cleavage activity of the hammerhead ribozyme induced by $Mg^{2+}$ was investigated using the solutions containing background nucleic acids, small phosphate and carboxylic acid compounds, and neutral polymers. Analysis of the substrate cleavage kinetics showed that the anionic compounds do not affect the ribozyme activity in $Mg^{2+}-saturated$ solutions and there is almost no effect of the $anion–Mg^{2+}$ complexes formed. On the other hand, the rate of substrate cleavage in $Mg^{2+}-unsaturated$ solutions was reduced under conditions of a high background of anionic compounds found in cells. The extent of the reduction was more with a greater net negative charge, caused by decreased amounts of $Mg^{2+}$ that could be used for the ribozyme reaction. It was remarkable that background DNA and RNA molecules having phosphodiester bonds reduced the cleavage rate as much as adenosine monophosphates having a charge of −2 when the effects of the same amount of phosphate groups were compared. Greater reductions in rates than those expected from the molecular charge were also observed in the background of fatty acids that form micelles. An addition of poly(ethylene glycol) to the solutions partially restored the ribozyme activity, suggesting a possible role of macromolecular crowding in counteracting the inhibitory effects of background anions on the ribozyme reaction. The results have biological and practical implications with respect to the effects of molecular environment on the efficiency of ion binding to RNA.
File Format	HTM / HTML
ISSN	09498257
Issue Number	6
Volume Number	20
e-ISSN	14321327
Journal	JBIC Journal of Biological Inorganic Chemistry
Language	English
Publisher	Springer
Publisher Date	2015-09-01
Publisher Place	Germany
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Biochemistry Anions Chemistry Rna, Catalytic Antagonists & Inhibitors Adenine Nucleotides Amino Acids Base Sequence Dextrans Fatty Acids Glucose-6-phosphate Kinetics Magnesium Polyethylene Glycols Rna Cleavage Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Biochemistry Inorganic Chemistry

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