Effects of background anionic compounds on the activity of the hammerhead ribozyme in Mg2+-unsaturated solutions

Verfasser / Beitragende:
[Shu-ichi Nakano, Yuichi Kitagawa, Daisuke Miyoshi, Naoki Sugimoto]
Ort, Verlag, Jahr:
2015
Enthalten in:
JBIC Journal of Biological Inorganic Chemistry, 20/6(2015-09-01), 1049-1058
Format:
Artikel (online)
ID: 605507031
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024 7 0 |a 10.1007/s00775-015-1286-y  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00775-015-1286-y 
245 0 0 |a Effects of background anionic compounds on the activity of the hammerhead ribozyme in Mg2+-unsaturated solutions  |h [Elektronische Daten]  |c [Shu-ichi Nakano, Yuichi Kitagawa, Daisuke Miyoshi, Naoki Sugimoto] 
520 3 |a 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 Mg2+ 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 Mg2+-saturated solutions and there is almost no effect of the anion-Mg2+ complexes formed. On the other hand, the rate of substrate cleavage in Mg2+-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 Mg2+ 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. 
540 |a SBIC, 2015 
690 7 |a Ribozyme  |2 nationallicence 
690 7 |a Metal ion binding  |2 nationallicence 
690 7 |a Macromolecular crowding  |2 nationallicence 
690 7 |a Poly(ethylene glycol)  |2 nationallicence 
700 1 |a Nakano  |D Shu-ichi  |u Department of Nanobiochemistry, Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, 7-1-20, Minatojima-minamimachi, Chuo-ku, 650-0047, Kobe, Japan  |4 aut 
700 1 |a Kitagawa  |D Yuichi  |u Department of Nanobiochemistry, Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, 7-1-20, Minatojima-minamimachi, Chuo-ku, 650-0047, Kobe, Japan  |4 aut 
700 1 |a Miyoshi  |D Daisuke  |u Department of Nanobiochemistry, Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, 7-1-20, Minatojima-minamimachi, Chuo-ku, 650-0047, Kobe, Japan  |4 aut 
700 1 |a Sugimoto  |D Naoki  |u Department of Nanobiochemistry, Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, 7-1-20, Minatojima-minamimachi, Chuo-ku, 650-0047, Kobe, Japan  |4 aut 
773 0 |t JBIC Journal of Biological Inorganic Chemistry  |d Springer Berlin Heidelberg  |g 20/6(2015-09-01), 1049-1058  |x 0949-8257  |q 20:6<1049  |1 2015  |2 20  |o 775 
856 4 0 |u https://doi.org/10.1007/s00775-015-1286-y  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00775-015-1286-y  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Nakano  |D Shu-ichi  |u Department of Nanobiochemistry, Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, 7-1-20, Minatojima-minamimachi, Chuo-ku, 650-0047, Kobe, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Kitagawa  |D Yuichi  |u Department of Nanobiochemistry, Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, 7-1-20, Minatojima-minamimachi, Chuo-ku, 650-0047, Kobe, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Miyoshi  |D Daisuke  |u Department of Nanobiochemistry, Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, 7-1-20, Minatojima-minamimachi, Chuo-ku, 650-0047, Kobe, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Sugimoto  |D Naoki  |u Department of Nanobiochemistry, Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, 7-1-20, Minatojima-minamimachi, Chuo-ku, 650-0047, Kobe, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t JBIC Journal of Biological Inorganic Chemistry  |d Springer Berlin Heidelberg  |g 20/6(2015-09-01), 1049-1058  |x 0949-8257  |q 20:6<1049  |1 2015  |2 20  |o 775