Probing tRNA interaction with biogenic polyamines

作者：Ouameur Amin Ahmed; Bourassa Philippe; Tajmir Riahi Heidar Ali^*

来源：RNA-A Publication of the RNA Society, 2010, 16(10): 1968-1979.

DOI：10.1261/rna.1994310

• tRNA
• polyamines
• preferential binding site
• binding constant
• hyperchromism
• hypochromism
• condensation
• tRNA stability
• FTIR spectroscopy
• affinity capillary electrophoresis (ACE)

Biogenic polyamines are found to modulate protein synthesis at different levels. This effect may be explained by the ability of polyamines to bind and influence the secondary structure of tRNA, mRNA, and rRNA. We report the interaction between tRNA and the three biogenic polyamines putrescine, spermidine, spermine, and cobalt(III) hexamine at physiological conditions, using FTIR spectroscopy, capillary electrophoresis, and molecular modeling. The results indicated that tRNA was stabilized at low biogenic polyamine concentration, as a consequence of polyamine interaction with the backbone phosphate group. The main tRNA reactive sites for biogenic polyamine at low concentration were guanine-N7/O6, uracil-O2/O4, adenine-N3, and 2'OH of the ribose. At high polyamine concentration, the interaction involves guanine-N7/O6, adenine-N7, uracil-O2 reactive sites, and the backbone phosphate group. The participation of the polycation primary amino group, in the interaction and the presence of the hydrophobic contact, are also shown. The binding affinity of biogenic polyamine to tRNA molecule was in the order of spermine > spermidine > putrescine with K(Spm) = 8.7 x 10(5) M(-1), K(Spd) = 6.1 x 105 M(-1), and K(Put) = 1.0 x 10(5) M(-1), which correlates with their positively charged amino group content. Hill analysis showed positive cooperativity for the biogenic polyamines and negative cooperativity for cobalt-hexamine. Cobalt(III) hexamine contains high-and low-affinity sites in tRNA with K(1) = 3.2 x 10(5) M(-1) and K(2) = 1.7 x 10(5) M(-1), that have been attributed to the interactions with guanine-N7 sites and the backbone PO(2) group, respectively. This mechanism of tRNA binding could explain the condensation phenomenon observed at high Co(III) content, as previously shown in the Co(III)-DNA complexes.

• 出版日期2010-10