Explanation by the double-metal-ion mechanism of catalysis for the differential metal ion effects on the cleavage rates of 5′-oxy and 5′-thio substrates by a hammerhead ribozyme
AUTOR(ES)
Zhou, De-Min
FONTE
The National Academy of Sciences of the USA
RESUMO
In a previous examination using natural all-RNA substrates that contained either a 5′-oxy or 5′-thio leaving group at the cleavage site, we demonstrated that (i) the attack by the 2′-oxygen at C17 on the phosphorus atom is the rate-limiting step only for the substrate that contains a 5′-thio group (R11S) and (ii) the departure of the 5′ leaving group is the rate-limiting step for the natural all-RNA substrate (R11O) in both nonenzymatic and hammerhead ribozyme-catalyzed reactions; the energy diagrams for these reactions were provided in our previous publication. In this report we found that the rate of cleavage of R11O by a hammerhead ribozyme was enhanced 14-fold when Mg2+ ions were replaced by Mn2+ ions, whereas the rate of cleavage of R11S was enhanced only 2.2-fold when Mg2+ ions were replaced by Mn2+ ions. This result appears to be exactly the opposite of that predicted from the direct coordination of the metal ion with the leaving 5′-oxygen, because a switch in metal ion specificity was not observed with the 5′-thio substrate. However, our quantitative analyses based on the previously provided energy diagram indicate that this result is in accord with the double-metal-ion mechanism of catalysis.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=24970Documentos Relacionados
- A two-metal ion mechanism operates in the hammerhead ribozyme-mediated cleavage of an RNA substrate
- Observations on catalysis by hammerhead ribozymes are consistent with a two-divalent-metal-ion mechanism
- A further investigation and reappraisal of the thio effect in the cleavage reaction catalyzed by a hammerhead ribozyme
- Effects of variations in length of hammerhead ribozyme antisense arms upon the cleavage of longer RNA substrates.
- Importance in catalysis of a magnesium ion with very low affinity for a hammerhead ribozyme