Effect of a quaternary pentamine on RNA stabilization and enzymatic methylation
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Armine Hayrapetyan
Abstract
Extreme thermophiles produce unusually long polyamines, including the linear caldopentamine (Cdp) and the branched pentamine tetrakis(3-aminopropyl)-ammonium (Taa), with the latter containing a central quaternary ammonium moiety. Here we compare the interaction of these two pentamines with RNA by studying the heat denaturation, electrophoretic behavior, and ability of tRNA to be methylated in vitro by purified tRNA methyltransferases under various salt conditions. At concentrations in the micromolar range, branched Taa causes a considerable increase in the melting temperature (Tm) of yeast tRNAPhe transcripts by >20°C, which is significantly greater than stabilization by the linear Cdp. In non-denaturing gel electrophoresis, strong and specific binding to Taa, but not to Cdp, was clearly observed for tRNAPhe. In both types of experiments, polyamines and monovalent metal ions competed for binding sites. Structural probing revealed no significant conformational changes in tRNA on Taa binding. In post-transcriptional in vitro methylation reactions, the formation of m2G/m22G by the methyltransferase Trm1p and of m1A by TrmIp were not affected or only slightly stimulated by polyamines. In contrast, Taa specifically inhibited Trm4p-dependent formation of m5C only in tRNAPhe, likely by occupying sites that are relevant to RNA recognition by the methyltransferase.
©2009 by Walter de Gruyter Berlin New York
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- Review
- Dimeric/oligomeric DNA methyltransferases: an unfinished story
- Minireview
- Induced human pluripotent stem cells: promises and open questions
- Genes and Nucleic Acids
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- Protein Structure and Function
- A soluble form of ammonia monooxygenase in Nitrosomonas europaea
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- A dual role of the N-terminal FQQI motif in GLUT4 trafficking
- Molecular Medicine
- Peritumoral administration of GPI-anchored TIMP-1 inhibits colon carcinoma growth in Rag-2 γ chain-deficient mice
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- The effect of endogenous preproneuropeptide Y leucine 7 to proline 7 polymorphism on growth and apoptosis in primary cultured HUVECs
- Cross talk between kinin and angiotensin II receptors in mouse abdominal aorta
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- Association between kallikrein-related peptidases (KLKs) and macroscopic indicators of semen analysis: their relation to sperm motility
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