Abstract
The binding of Mg(2+)/Mn(2+) to acceptor stem microhelices as minimal models for precursor-tRNA(Gly) is demonstrated by NMR spectroscopy. From the evaluation of COSY and NOESY spectra, binding sites for Mg(2+)/Mn(2+) can be inferred. In particular, one binding site exists near the ribose moiety of nucleotide -1 at the position of cleavage by RNase P. From comparison with a variant possessing a deoxynucleotide at this position, it is concluded that the 2'-OH group of this nucleotide is indispensable for coordinating the divalent metal ion. Hence, this catalytically important metal ion is "pre-bound" to the precursor-tRNA before complexation with RNase P.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Base Pairing
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Base Sequence
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Binding Sites
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Cations, Divalent / metabolism
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Endoribonucleases / chemistry
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Endoribonucleases / genetics
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Endoribonucleases / metabolism*
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Escherichia coli / enzymology*
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Escherichia coli / genetics
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Escherichia coli Proteins*
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Magnesium / metabolism*
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Manganese / metabolism*
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Nuclear Magnetic Resonance, Biomolecular
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Protein Binding
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RNA Precursors / chemistry
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RNA Precursors / genetics
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RNA Precursors / metabolism*
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RNA, Catalytic / chemistry
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RNA, Catalytic / genetics
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RNA, Catalytic / metabolism*
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RNA, Transfer / chemistry
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RNA, Transfer / genetics
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RNA, Transfer / metabolism*
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Ribonuclease P
Substances
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Cations, Divalent
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Escherichia coli Proteins
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RNA Precursors
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RNA, Catalytic
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Manganese
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RNA, Transfer
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Endoribonucleases
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Ribonuclease P
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ribonuclease P, E coli
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Magnesium