Biver Tarita, Busto Natalia, García Begoña, Leal José M, Menichetti Luisa, Secco Fernando, Venturini Marcella
Dipartimento di Chimica e Chimica Industriale, Università di Pisa, Via Moruzzi 13, 56124 Pisa, Italy.
Departamento de Química, Universidad de Burgos, Plaza Misael Bañuelos s/n, 09001 Burgos, Spain.
J Inorg Biochem. 2015 Oct;151:115-22. doi: 10.1016/j.jinorgbio.2015.05.001. Epub 2015 May 9.
The ability of magnesium(II) and nickel(II) to induce dramatic conformational changes in the synthetic RNA poly(rA)poly(rU) has been investigated. Kinetic experiments, spectrofluorometric titrations, melting experiments and DSC measurements contribute in shedding light on a complex behaviour where the action of metal ions (Na(+), Mg(2+), Ni(2+)), in synergism with other operators as the intercalating dye coralyne and temperature, all concur in stabilising a peculiar RNA form. Mg(2+) and Ni(2+) (M) bind rapidly and almost quantitatively to the duplex (AU) to give a RNA/metal ion complex (AUM). Then, by the union of two AUM units, an unstable tetra-aggregate (UAUA(M2)) is formed which, in the presence of a relatively modest excess of metal, evolves to the UAUM triplex by releasing a single AM strand. On the other hand, under conditions of high metal content, the UAUA(M2) intermediate rearranges to give a more stable tetra-aggregate (UAUA(M2)). As concerns the role of coralyne (D), it is found that D strongly interacts with UAUA(M2). Also, in the presence of coralyne, the ability of divalent ions to promote the transition of AUD into UAUD is enhanced, according to the efficiency sequence [Ni(2+)]≫[Mg(2+)]≫[Na(+)].
已对镁(II)和镍(II)在合成RNA聚(rA)聚(rU)中诱导显著构象变化的能力进行了研究。动力学实验、荧光光谱滴定、熔解实验和差示扫描量热法测量有助于揭示一种复杂行为,其中金属离子(Na⁺、Mg²⁺、Ni²⁺)与嵌入染料珊瑚灵和温度等其他因素协同作用,共同稳定一种特殊的RNA形式。Mg²⁺和Ni²⁺(M)迅速且几乎定量地与双链体(AU)结合,形成RNA/金属离子复合物(AUM)。然后,通过两个AUM单元的结合,形成一个不稳定的四聚体(UAUA(M₂)*),在相对适量过量的金属存在下,通过释放一条AM链演变为UAUM三链体。另一方面,在高金属含量条件下,UAUA(M₂)*中间体重新排列形成更稳定的四聚体(UAUA(M₂))。关于珊瑚灵(D)的作用,发现D与UAUA(M₂)强烈相互作用。此外,在存在珊瑚灵的情况下,二价离子促进AUD转变为UAUD的能力增强,遵循效率顺序[Ni²⁺]≫[Mg²⁺]≫[Na⁺]。