Synthetic riboswitches for the analysis of tRNA processing by eukaryotic RNase P enzymes.

Autor:	Ender A; Institute for Biochemistry, Leipzig University, 04103 Leipzig, Germany., Grafl N; Institute for Biochemistry, Leipzig University, 04103 Leipzig, Germany., Kolberg T; Institute for Biochemistry, Leipzig University, 04103 Leipzig, Germany., Findeiß S; Bioinformatics Group, Department of Computer Science and Interdisciplinary Center for Bioinformatics, Leipzig University, 04107 Leipzig, Germany., Stadler PF; Bioinformatics Group, Department of Computer Science and Interdisciplinary Center for Bioinformatics, Leipzig University, 04107 Leipzig, Germany.; Max Planck Institute for Mathematics in the Science, 04103 Leipzig, Germany.; Institute for Theoretical Chemistry, University of Vienna, A-1090 Vienna, Austria.; Santa Fe Institute, Santa Fe, New Mexico 87501, USA., Mörl M; Institute for Biochemistry, Leipzig University, 04103 Leipzig, Germany.
Jazyk:	angličtina
Zdroj:	RNA (New York, N.Y.) [RNA] 2022 Apr; Vol. 28 (4), pp. 551-567. Date of Electronic Publication: 2022 Jan 12.
DOI:	10.1261/rna.078814.121
Abstrakt:	Removal of the 5'-leader region is an essential step in the maturation of tRNA molecules in all domains of life. This reaction is catalyzed by various RNase P activities, ranging from ribonucleoproteins with ribozyme activity to protein-only forms. In Escherichia coli , the efficiency of RNase P-mediated cleavage can be controlled by computationally designed riboswitch elements in a ligand-dependent way, where the 5'-leader sequence of a tRNA precursor is either sequestered in a hairpin structure or presented as a single-stranded region accessible for maturation. In the presented work, the regulatory potential of such artificial constructs is tested on different forms of eukaryotic RNase P enzymes-two protein-only RNase P enzymes (PRORP1 and PRORP2) from Arabidopsis thaliana and the ribonucleoprotein of Homo sapiens The PRORP enzymes were analyzed in vitro as well as in vivo in a bacterial RNase P complementation system. We also tested in HEK293T cells whether the riboswitches remain functional with human nuclear RNase P. While the regulatory principle of the synthetic riboswitches applies for all tested RNase P enzymes, the results also show differences in the substrate requirements of the individual enzyme versions. Hence, such designed RNase P riboswitches represent a novel tool to investigate the impact of the structural composition of the 5'-leader on substrate recognition by different types of RNase P enzymes. (© 2022 Ender et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)
Databáze:	MEDLINE
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