Metal-Ion-Dependent Catalysis and Specificity of CCA-Adding Enzymes: A Comparison of Two Classes
| Autor: | Ya-Ming Hou, Lindsey Ingerman, Shan-Qing Gu, Haiping Zhou |
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| Rok vydání: | 2005 |
| Předmět: |
Cations
Divalent Stereochemistry Metal ions in aqueous solution Molecular Sequence Data Biochemistry Catalysis Substrate Specificity Sulfolobus chemistry.chemical_compound Escherichia coli Nucleotide RNA Transfer Val Polymerase chemistry.chemical_classification Base Sequence biology RNA RNA Nucleotidyltransferases biology.organism_classification Enzyme chemistry Metals Transfer RNA biology.protein DNA |
| Zdroj: | Biochemistry. 44:12849-12859 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi0509402 |
| Popis: | The CCA-adding enzymes [ATP(CTP):tRNA nucleotidyl transferases] catalyze synthesis of the conserved and essential CCA sequence to the tRNA 3' end. These enzymes are divided into two classes of distinct structures that differ in the overall orientation of the head to tail domains. However, the catalytic core of the two classes is conserved and contains three carboxylates in a geometry commonly found in DNA and RNA polymerases that use the two-metal-ion mechanism for phosphoryl transfer. Two important aspects of the two-metal-ion mechanism are tested here for CCA enzymes: the dependence on metal ions for catalysis and for specificity of nucleotide addition. Using the archaeal Sulfolobus shibabae enzyme as an example of the class I, and the bacterial Escherichia coli enzyme as an example of the class II, we show that both enzymes depend on metal ions for catalysis, and that both use primarily Mg2+ and Mn2+ as the "productive" metal ions, but several other metal ions such as Ca2+ as the "nonproductive" metal ions. Of the two productive metal ions, Mg2+ specifically promotes synthesis of the correct CCA, whereas Mn2+ preferentially accelerates synthesis of the noncognate CCC and poly(C). Thus, despite evolution of structural diversity of two classes, both classes use metal ions to determine catalysis and specificity. These results provide critical insights into the catalytic mechanism of CCA synthesis to allow the two classes to be related to each other, and to members of the larger family of DNA and RNA polymerases. |
| Databáze: | OpenAIRE |
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