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The Plant tRNA 3‘ Processing Enzyme Has a Broad Substrate Spectrum

Schiffer, Steffen and Helm, Mark and Théobald-Dietrich, Anne and Giegé, Richard and Marchfelder, Anita (2001) The Plant tRNA 3‘ Processing Enzyme Has a Broad Substrate Spectrum. Biochemistry, 40 (28). pp. 8264-8272. ISSN 0006-2960. doi:10.1021/bi0101953.

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To elucidate the minimal substrate for the plant nuclear tRNA 3‘ processing enzyme, we synthesized a set of tRNA variants, which were subsequently incubated with the nuclear tRNA 3‘ processing enzyme. Our experiments show that the minimal substrate for the nuclear RNase Z consists of the acceptor stem and T arm. The broad substrate spectrum of the nuclear RNase Z raises the possibility that this enzyme might have additional functions in the nucleus besides tRNA 3‘ processing. Incubation of tRNA variants with the plant mitochondrial enzyme revealed that the organellar counterpart of the nuclear enzyme has a much narrower substrate spectrum. The mitochondrial RNase Z only tolerates deletion of anticodon and variable arms and only with a drastic reduction in cleavage efficiency, indicating that the mitochondrial activity can only cleave bona fide tRNA substrates efficiently. Both enzymes prefer precursors containing short 3‘ trailers over extended 3‘ additional sequences. Determination of cleavage sites showed that the cleavage site is not shifted in any of the tRNA variant precursors.

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Additional Information:© 2001 American Chemical Society. Received 30 January 2001. Published online 21 June 2001. Published in print 1 July 2001. Work presented here was funded by the Anfangsförderung Universität Ulm, the Deutsche Forschungsgemeinschaft, and the Fonds der Chemischen Industrie. We thank Dr. Axel Brennicke for critical reading of the manuscript, Drs. U. Gerischer and P. Dürre for use of the FujiFilm Image Reader, and Dr. A. Kunzmann for the highly purified mitochondrial RNase Z. We appreciate the expert technical assistance of Elli Bruckbauer and Claudia Gautsch.
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Anfangsförderung Universität UlmUNSPECIFIED
Deutsche Forschungsgemeinschaft (DFG)UNSPECIFIED
Fonds der Chemischen IndustrieUNSPECIFIED
Issue or Number:28
Record Number:CaltechAUTHORS:20170131-102251011
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Official Citation:The Plant tRNA 3‘ Processing Enzyme Has a Broad Substrate Spectrum Steffen Schiffer, Mark Helm, Anne Théobald-Dietrich, Richard Giegé, and Anita Marchfelder Biochemistry 2001 40 (28), 8264-8272 DOI: 10.1021/bi0101953
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:73874
Deposited By: Ruth Sustaita
Deposited On:31 Jan 2017 19:52
Last Modified:11 Nov 2021 05:23

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